Spectrum Diagnostics Products List 2014-2015

Product Inserts 2014
www.spectrum-diagnostics.com
The creative
Approach
to Bioscience
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Obour city industrial area. block 20008 piece 19 A. Cairo. Egypt.
Tel: +2 02 4665 1848 Fax: +2 02 4665 1847
E-mail: info@spectrum-diagnostics.com
Contents E- Infectious immunology, Bacteriology 2 34
CREATINE KINASE MB 86 Rheumatoid Factor (RF) 170
g - Glutamyltransferase-(gGT)-Liquizyme (9+1) 88
D- turbidimetric immunoassay 173 Brucella A, Brucella M, Brucella Combo 236
A- Clinical Chemistry 7 g - Glutamyltransferase-(gGT)-Liquizyme (1+1) 90
Antistreptolysin O (ASO) Immuno-Turbidimetry 174 Widal Test (Salmonella Ab) 238
Aspartate aminotransferase (AST/GOT)-Liquizyme (4+1) 92 RPR SYPHILIS TEST 240
i-Substrates 8 Antistreptolysin O (ASO) Turbi Latex 176
Aspartate aminotransferase (AST/GOT)-Liquizyme (1 + 1) 94 Rose Bengal Brucella Antigen 242
Albumin - BCG 10 ASO Standard Super High 178
Aspartate aminotransferase (AST/GOT)-Colorimetric 96 TOXO LATEX KIT 244
Bilirubin (TOTAL AND DIRECT) 12 ASO Control 179
Aspartate aminotransferase 98
Pediatric Total BILIRUBIN 14 b2 -MICROGLOBULIN (b2 -m) 180 F- Hematology, Haemostasis Reagents 2 47
Alanine aminotransferase (ALT/GPT)-Liquizyme (4+1) 100
TOTAL Bilirubin 16 b2 -MICROGLOBULIN Mono-Reagent Procedure 182 PT Reagent (SP-NORMOPLASTIN) 248
Alanine aminotransferase (ALT/GPT)-Liquizyme (1 + 1) 102
Bilirubin (DIRECT) 18 C REACTIVE PROTEIN (CRP) Immuno-Turbidimetry 184 NORMAL AND ABNORMAL PLASMA CONTROLS FOR COAGULATION ASSAYS 250
Alanine aminotransferase (ALT/GPT) - Colorimetric 104
Bilirubin (TOTAL AND DIRECT) 20 C REACTIVE PROTEIN (CRP) Turbi Latext 186 SP-Ultraplastin PT Reagent ISI 1.05 252
Alanine aminotransferase (ALT/GPT) - Ultimate Single Reagent 106
Cholesterol – Liquizyme 22 Ceruloplasmin Immuno-Turbidimetry 188 APTT Reagent (SP- UNICELIN) 254
Glucose-6-Phosphate dehydrogenase (G-6-PDH) 108
HDL CHOLESTEROL - Precipitant 24 CRP Standard Super High 189
Glucose-6-Phosphate 110 G-blood group 256
HDL CHOLESTEROL 26 CRP Control 190
Lactate dehydrogenase (LDH)-Liquizyme (4+1) 112 Spectrum Anti-A, Anti-B,Anti-AB Monoclonal (IgM) 258
LDL CHOLESTEROL 28 CRP/hs(C- Reactive protein) 192
Lactate dehydrogenase (LDH)-Liquizyme (1+1) 114 Anti-D (Rho) Plus Monoclonal IgM & IgG 260
Creatinine – Jaffè 30 CYSTATIN-C Immunoturbidimetry 194
LIPASE-LS COLORIMETRIC (DGMRE) 116 ANTI-HUMAN GLOBULIN SERUM (COOMBS) 262
Creatinine – Jaffè 32 FERRITIN Turbi Latex 196
BOVINE SERUM ALBUMIN (BSA) 264
Creatinine - Colorimetric 34 iii-Electrolytes 119 FIBRINOGEN Immuno - Turbidimetry 198
Low Ionic Strength (LISS) Solution 266
GLUCOSE - Liquizyme 36 Ammonia II (GLDH - UV - Test) 120 Fibrinogen Standard 200
GLYCOSYLATED HEMOGLOBIN (GHb) 38 Ammonia – Liquizyme Single Reagent 122 HAPTOGLOBIN (HAP) Immuno Turbidimetry 202 H-Dry chemistry 269
Calcium O-CPC 124 Hemoglobin A1c Standard Set 203 CMV IgG/IgM Rapid Test - Cassette 270
Direct Enzymatic HbA1c 40
Calcium Arsenazo III (Single Reagent) 126 Hemoglobin A1c (HbA1c) Immuno - Turbidimetry 204 H. pylori Ab Test Device (Serum / Plasma / Whole Blood) 272
Haemoglobin(Single Reagent) 42
Chloride Single Reagent 128 IMMUNOGLOBULIN E (IgE) 206 H. pylori Ag Test Device (Fecal Specimen) 274
Haemoglobin 44
Carbon Dioxide (CO2) (Colorimetric PEPC) 130 Immunology Control High 208 HAV IgM Rapid Test-Cassette (Serum / Plasma) 276
Haemoglobin (Ready-To-Use) 46
Copper 132 Immunology Control Low 209 HBsAg Hepatitis B Surface Antigen Test Device 278
Lactate – Liquizyme 48
Total Iron Chromazurol B Single Reagent 134 Kappa light chain serum kit (KAP) 210 HBsAg/HCV Ab Rapid Test - Cassette 280
Micrototal Protein (MT-P) 50
Total Iron and Total Iron Binding Capacity Reagent Set 136 Kappa light chain urine kit (KAP) 211 HCG One Step Pregnancy Test Device (Urine/Serum) 284
PYRUVATE 52
Total Iron Binding Capacity (TIBC) 138 Lambda light chain serum kit (LAM) 212 HCV Ab Hepatitis C Virus Ab Test Device (Serum/Plasma) 286
Total bile acids (TBA) 54
Total Iron - Ferrozine 140 Lambda light chain urine kit (LAM) 213 HIV Ab 1/2/O 288
Total protein 56
MAGNESIUM Xylidyl Blue Monoreagnt 142 LIPOPROTEIN (a) [Lp(a)] Mono-Reagent Procedure 214 Malaria Antigen Test (Whole Blood) 290
Triglycerides – Liquizyme 58
Phosphorus, Inorganic 144 Microalbumin Standard 216 S.typhi-S.paratyphi “A” Direct Antigen Detection 292
Urea/BUN - Liquizyme 60
INORGNIC PHOSPHORUS 146 MICROALBUMIN (MAU) Immuno Turbidimetry 218 Syphilis Ab Combo Rapid Test 294
Urea/BUN - LS 62
Potassium 148 MICROALBUMIN (MAU) Turbi Latex 220 ToRCH 296
Urea/BUN – Liquizyme (UV) 64
Sodium Single Reagent 150 Microalbumin Control set 222 Toxo IgG/IgM Rapid Test- Cassette 298
Urea/BUN – (UV) 66
ZINC (Colorimetric Test with 5-Brom-PAPS) 152 Protein standard High 223 Troponin-I 300
Uric acid - Liquizyme 68
Procalcitonin Assay 224 Troponin-I 302
ii-Enzymes 71 B-Quality control 155
RF Control 226 Typhoid IgG/IgM Rapid Test - Cassette 304
Acid Phosphatase (ACP) 72 SP-NORMOTROL 156
RF Standard Super High 227 URI-TRAK 306
Alkaline phosphatase (ALP) 74 SP-PATHOTROL 158
RHEUMATOID FACTOR (RF) ImmunoTurbidimetry 3rd Generation 228 URI-TRAK-10 308
Alkaline phosphatase (ALP) 76 SP-MULTICAL 160
RHEUMATOID FACTOR (RF) Turbi Latex 230
ALKALINE PHOSPHATASE 78 HbA1c Calibrator 162
Specific Protein Control 232
Alkaline Phosphatase - Colorimetric 80 C-immunochemistry 165 Specific Protein standard 233
Alpha Amylase 82 Antistreptolysin O Titre (ASOT) 166
Creatine Kinase (CK) 84 C Reactive Protein (CRP) 168
A- Clinical Chemistry
6 7
i-Substrates
8 9
Albumin - BCG SYMBOLS IN PRODUCT LABELLING
Methods Comparison References
1. Doumas BT,Watson WA, Biggs HG. Albumin standard and the
(Acetate Buffer) EC REP Authorised Representative Temperature Limitation A comparison between Spectrum Diagnostics Albumin reagent and
measurement of serum albumin with bromocresol green Clin
IVD For in-vitro diagnostic use Use by/Expiration Date a commercial reagent of the same methodology was performed on
20 human sera. A correlation of 0.97 was obtained. Chim Acta. 1971;31:87-96.
REF: 211 001 (2x100 ml) 200 test LOT Batch Code/Lot number CAUTION. Consult instructions
REF: 211 002 (4x100 ml) 400 test REF Catalogue Number for use 2. Grant GH, Silverman LM, Christenson RH. Amino acids and
REF: 211 003 (2x500 ml) 1000 test Consult instructions for use Manufactured by Sensitivity proteins. In:Tietz NW, ed. Fundamentals of Clinical Chemistry.
REF: 211 004 (2x250 ml) 500 test 3rd ed. Philadelphia:WB Saunders;1987:291 345.
When run as recommended, the minimum detection limit of this
Intended Use Specimen Collection and Preservation assay is 1.0 g/dL. 3. Tietz NW, ed. Clinical Guide to laboratory tests. 2nd ed.
Philadelphia: WB Saunders; 1990:26-29.
Spectrum Diagnostics albumin reagent is intended for the in- v i t r o The only acceptable anticoagulats are heparin and EDTA. Use Linearity
quantitative,diagnostic determination of albumin in human serum on preferably fresh serum, Serum should be separated immediately
both automated and manual systems. from the clot. The biological half-life of albumin in blood is 3 weeks. The reaction is linear up to an albumin concentration of 7.0 g/dL;
o o
Stability: 1 day at 15 – 25 C; 4 weeks at 4 – 8 C; specimens showing higher concentration should be diluted 1+1 with ORDERING INFORMATION
o
6 months at -20 C
Background physiological saline and repeat the assay (result × 2).
CATALOG NO. QUANTITY
Albumin is the major serum protein in normal individuals. It maintains System Parameters Interfering Substances 211 001 2 x 100 ml
the plasma colloidal osmotic pressure, binds and solubilizes many Serum, plasma 211 002 4 x 100 ml
compounds such as calcium and bilirubin. Elevated serum albumin Wavelength 623 nm ( or 578 nm ) 211 003 2 x 500 ml
levels are usually the result of dehydration. Hyperalbuminemia is Optical path 1 cm 211 004 2 x 250 ml
Haemolysis
of little diagnostic significance. Hypoalbuminemia is very common Assay type End-point
A haemoglobin level of 800 mg/dL results in 13 % positive bias.
in many diseases including malabsorption, liver diseases. kidney Direction Increase
diseases, severe burns, infections, cancer and some genetic Sample : Reagent Ratio 1 : 100
Icterus
abnormalities. In severe hypoalbuminemia (less than 2.5 g/dL), the e.g.: Reagent volume 1 ml
No significant interference up to a bilirubin level of 40 mg/dL.
low plasma oncotic pressure allows water to move out of the blood Sample volume 10 ml
o
capillaries into the tissues causing edema. Temperature 20 – 25 C
o Lipemia
Incubation time 5 minutes at 20–25 C
No significant interference up to an intralipid level of 1000 mg/dL.
Zero adjustment Reagent Blank
Method
Sensitivity 1 g/dL
Linearity 7 g/dL Expected Values
Modified bromocresol green colorimetric method.
Adults
Assay Principle Procedure
18 – 60 y 3.5 – 5.5 g/dL (35 – 50 g/L)
Measurement of albumin is based on its binding to the indicator dye Blank Standard specimen >60 y 3.4 – 4.8 g/dL (34 – 48 g/L)
bromocresol green (BCG) in pH 4.1 to form a blue-green colored
complex. The intensity of the blue-green color is directly proportional Reagent (R) 1 ml 1 ml 1 ml
Childern
to the concentration of albumin in the sample. It is determined by Standard ------- 10 ml -------
monitoring the increase in absorbance at 623 nm, or 578 nm. 14-18 y 3.2-4.5 g/dL (32-45 g/L)
Specimen ------- ------- 10 ml
4d-14 y 3.8-5.4 g/dL (38-54 g/L)
pH 4.1
Albumin + BCG Albumin-BCG Complex o
Mix , incubate for approximately 5 minutes at 20-25 C.
Newborns
Measure absorbance of specimen (Aspecimen) and standard
Reagents (Astandard) against reagent blank within 60 minutes.
0-4 day 2.8-4.4 g/dL (28-44 g/L)
Standard albumin
Calculation Spectrum Diagnostics does not interpret the results of a
4.0 g/dL.
Aspecimen clinical laboratory procedure ; interpretation of the results is
Albumin concentration (g/dL) = x4 considered the responsibility of qualified medical personnel.
Reagent (R) Astandard All indications of clinical significance are supported by
Acetate Buffer 100 mmol/L
literature references.
Bromocresol green 0.27 mmol/L
Detergent Quality Control
For further information, refer to the Albumin reagent material safety Normal & abnormal commercial control serum of known
concentrations should be analyzed with each run.
Analytical Range
data sheet.
1.0 – 7.0 g/dL.
Performance Characteristics
Precautions and Warnings
Precision
Within run (Repeatiblity) Waste Disposal
Do not ingest or inhalate. In case of contact with eyes or skin; rinse
immediately with plenty of soap and water. In case of severe injuries; Level 1 Level 2 This product is made to be used in professional laboratories.
seek medical advice immediately. Please consult local regulations for a correct waste disposal.
n 20 20
S56: dispose of this material and its container at hazardous or
Mean (g/dL) 3.28 4.78 special waste collection point.
SD 0.8 0.12 S57: use appropriate container to avoid environmental
Reagent Preparation, Storage and Stability contamination.
CV% 2.66 2.68
S61: avoid release in environment. refer to special instructions/
Spectrum albumin reagents are supplied ready-to-use and stable safety data sheets.
up to the expiry date labeled on the bottles. The reagents are stable
o
at 2 – 8 C. Run to run (Reproducibility)
Level 1 Level 2
Deterioration n 20 20
Do not use the Spectrum albumin regents if precipitate forms. Mean (g/dL) 3.4 4.9
Failure to recover control values within the assigned range may SD 0.9 0.14
be an indication of reagent deterioration. Specimen Collection and
Preservation. CV% 3.1 2.9
10 11
Bilirubin (TOTAL AND DIRECT) SYMBOLS IN PRODUCT LABELLING
Direct Bilirubin Lipemia
Lipemic specimens interfere with the test.
Sample blank Sample
Jendrassik Grof EC REP Authorised Representative Use by/Expiration Date
Reagent 1 200 ml 200 ml Drugs
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions
LOT Batch Code/Lot number for use Reagent 2 ----- 1 drop Theophyllin and propranolol may cause artificially low total bilirubin
REF: 222 001 (255ml) 100 Test REF: 222 002 (750ml) 300 Test levels.
REF Catalogue Number Manufactured by
Saline 0.9% NaCl 2.0 ml 2.0 ml
Consult instructions for use (C) - Corrosive
R1 Sulphanilic Acid 1 x 45 ml R1 Sulphanilic Acid 2 x 65 ml
Temperature Limitation Sample 200 ml 200 ml Expected Values
R2 Nitrite 1 x 10 ml R2 Nitrite 2 x 15 ml o
Mix and incubate for exactly 5 minutes at 20 – 25 C. Measure Total Bilirubin
R3 Caffeine 1 x 100 ml R3 Caffeine 3 x 100 ml absorbance of sample (Asample) against sample blank at 546 nm Adults and infants>1 month < 0.2-1.0 mg/dL ( 3.4-17 mmol/l)
R4 Tartarate 1 x 100 ml R4 Tartarate 3 x 100 ml Reagent 4 contains caustic material. (530 - 560 nm). Newborns premature (3-5 d) 10-14 mg/dL (171-239 mmol/l)
Corrosive (C) Calculation Newborns:

Intended Use R35 Causes severe burns. Total bilirubin = ASample x 10.8 (3-5 d) 4.0 - 8.0 mg/dL ( 68-137 mmol/l)
R41 Risk of serious damage to eyes. Direct bilirubin = ASample x 14.4 (<48 h) 6.0 - 10.0 mg/dL (103-171 mmol/l)
Spectrum Diagnostics bilirubin reagent is intended for the in-vitro
S26 In case of contact with eyes, rinse immediately with (<24 h) 2.0-6.0 mg/dL (34-103 mmol/l)
quantitative, diagnostic determination of bilirubin in human serum
plenty of water and seek medical advice.
on both automated and manual systems. Quality Control
S28 After contact with skin, wash immediately with plenty of Direct Bilirubin 0 – 0.3 mg/dL (0 – 51 mmol/L)
Normal & abnormal commercial control serum of known
soap and water.
Background concentrations should be analyzed with each run.
For further information, refer to the •Bilirubin reagent material safety
data sheet.
The average level of the bilirubin produced in humans from different Performance Characteristics Spectrum Diagnostics does not interpret the results of a
sources ranges between 250 to 300 mg/day, of which 85% is derived Precision clinical laboratory procedure; interpretation of the results is
Precautions and Warnings considered the responsibility of qualified medical personnel.
from the heme moiety of the haemoglobin released from senescent
erythrocytes that are destroyed in the reticuloendothelial system. All indications of clinical significance are supported by
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Within run (Repeatiblity)
The remaining 15 % is produced from erythrocytes destroyed in the literature references.
immediately with plenty of soap and water. In case of severe injuries; Total Direct
bone marrow and from catabolism of other heme containing proteins
seek medical advice immediately.
such as cytochromes and myoglobin. Level 1 Level 2 Level 1 Level 2
After it is produced in the peripheral tissues, bilirubin is transported
to the liver in association with albumin. In the liver, bilirubin is Reagent Preparation, Storage and Stability n 20 20 20 20 Analytical Range
conjugated with glucuronic acid for solubilization and subsequent Mean (mg/dL) 0.79 4.37 0.299 0.77 Total bilirubin : 0.1 – 30 mg/dL ( 1.7 – 513 mmol/L)
transport through the bile duct and elimination via the digestive Spectrum bilirubin reagents are supplied ready-to-use and stable Direct bilirubin : 0.1 – 10 mg/dL ( 1.7 – 171 mmol/L)
up to the expiry date labeled on the bottles when stored at room SD 0.016 0.18 0.016 0.057
tract. Disease or conditions which, through hemolytic processes,
produce bilirubin faster than the liver can metabolize it, cause the temperature. CV% 2.13 4.12 5.41 7.4 Waste Disposal
levels of unconjugated (indirect) bilirubin to increase in the circulation. This product is made to be used in professional laboratories.
Bile duct obstruction or damage to hepatocellular structure causes Deterioration Run to run (Reproducibility) Please consult local regulations for a correct waste disposal.
increases in the levels of both conjugated (direct) and unconjugated S56: dispose of this material and its container at hazardous or
Do not use the Spectrum bilirubin reagents if precipitate forms. Total Direct special waste collection point.
(indirect) bilirubin in the circulation.
Failure to recover control values within the assigned range may be Level 1 Level 2 Level 1 Level 2 S57: use appropriate container to avoid environmental
Method an indication of reagent deterioration. contamination.
n 20 20 20 20
Colorimetric Diazo method. S61: avoid release in environment. refer to special instructions/
Specimen Collection and Preservation Mean (mg/dL) 0.82 4.52 0.32 0.82 safety data sheets.
Assay Principle SD 0.02 0.27 0.023 0.062
The total bilirubin concentration is determined in presence of Avoide exposure of the specimen to light. If plasma is used, only References
heparin and oxalate plasma are suitable. Other anticoagulants CV% 2.24 4.21 5.57 8.1
caffeine by the reaction with diazotized sulphanilic acid to produce 1. Balistreri WF, Shaw LM. Liver function. In: Tietz NW, ed.
an intensely colored diazo dye (560-600 nm). The intensity of color of should not be used. The average half-life of total bilirubin and direct
bilirubin in serum is 17 days and few hours respectively. Methods Comparison Fundamentals of clinical chemistry.3 rd ed. Philadelphia:WB
this dye formed is proportional to the concentration of total bilirubin. Saunders: 1987:729-761.
Direct bilirubin is determined in absence of caffeine by the direct
Stability: A comparison between Spectrum Diagnostics Bilirubin and a 2. Malloy HT, Evelyn KA. The determination of bilirubin with the
reaction with diazotized sulphanilic acid to form red-colored
o o o commercial reagent of the same methodology was performed on 20 photoelectric colorimetric method. J Biol Chem.1937:119:481
azobilirubin, the color intenisity of which measured at 546 nm is -20 C 4–8 C 20 – 25 C
human sera. A correlation of 0.975 was obtained. - 490.
proportional to the concentration of the direct bilirubin in the sample.
Total 6 months 7 days 1 day
HCL Sensitivity 3. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
Sulfanilic acid + NaNO2 Diazotized sulfanilic acid Direct 6 months 7 days 2 days
When run as recommended, the sensitivity of this assay is 0.1 mg/ ed.Philadephia: WB saunders; 1995:268-273.
pH 1.4 dL (1.7 mmol/L) for both total and direct bilirubin.
Bilirubin + Diazotized sulfanilic acid Azobilirubin Procedure
Total Bilirubin
Reagents Linearity
Sample blank Sample The reaction is linear up to a total bilirubin concentration of 30 mg/ ORDERING INFORMATION
Reagent 1 (R1) Reagent 1 200 ml 200 ml dL (513 mmol/L) and a direct bilirubin concentration of 10 mg/dL
(171 mmol/L). Specimens showing higher concentration should be CATALOG NO. QUANTITY
Sulfanilic acid 31.0 mmol/l Reagent 2 ----- 1 drop
HCL 0.20 N diluted 1+4 with physiological saline and repeat the assay (result×5). 222 001 100 test
Reagent 3 1.0 ml 1.0 ml 222 002 300 test
Reagent 2 (R2) Sample 200 ml 200 ml
Interfering substances
Sodium nitrite 28.0 mmol/l o
Serum, plasma
Mix and incubate for 10 minutes at 20 – 25 C. then add;
Reagent 3 (R3) Reagent 4 1.0 ml 1.0 ml Haemolysis
Caffeine 0.28 mol/l Avoid haemolysis since it interferes with the test.
Mix and incubate for 5 minutes at 20 – 25 oC. Measure absorbance
Sodium benzoate 0.55 mol/l
of sample (Asample) against sample blank at 578 nm(560 - 600
nm) The color intensity is stable for 30 minutes.
Reagent 4 (R4)
Tartarate 0.99 mol/l
Sodium hydroxide 2.0 N
12 13
Pediatric Total BILIRUBIN SYMBOLS IN PRODUCT LABELLING
Calculation
DA= A sample - A sample blank Waste Disposal
EC REP Authorised Representative Temperature Limitation Total Bilirubin Factor = 56 This product is made to be used in professional laboratories.
IVD For in-vitro diagnostic use Use by/Expiration Date Serum Total Bilirubin conc. ( mg/dL) = DA x 56 Please consult local regulations for a correct waste disposal.
LOT Batch Code/Lot number CAUTION. Consult instructions
REF: 221 001 ( 2 x 50 ml) 100 Test Conversion Factor = mg/dl x 17.1 = mmol/lQuality Control S56: dispose of this material and its container at hazardous or
REF Catalogue Number for use
REF: 221 002 ( 2 x 100 ml) 200 Test Normal & abnormal commercial control serum of known special waste collection point.
Consult instructions for use Manufactured by
concentrations should be analyzed with each run. S57: use appropriate container to avoid environmental
contamination.
Methods Comparison S61: avoid release in environment. refer to special instructions/
Intended Use Precautions and Warnings A comparison between Spectrum Diagnostics Total Bilirubin reagent safety data sheets.
Spectrum Diagnostics Total Bilirubin single reagent is intended for Do not ingest or inhalate. In case of contact with eyes or skin; rinse and a commercial reagent of the same methodology was performed
in-vitro quantitative, diagnostic determination of Total Bilirubin in immediately with plenty of soap and water. In case of severe injuries; on 20 human sera. A correlation of 0.978 was obtained. References
human serum on both manual and automated systems. seek medical advice immediately.
1. Balistreri WF, Shaw LM. Liver function. In: Tietz NW, ed.
Sensitivity Fundamentals of clinical chemistry.3 rd ed. Philadelphia:WB
Background Reagent Preparation, Storage and Stability When run as recommended, the minimum detection limit of the Saunders: 1987:729-761.
The average level of the bilirubin produced in humans from different The reagents are supplied ready to use. assay is 0.1 mg/dl (1.71 mmol/l)
o o
2. Malloy HT, Evelyn KA. The determination of bilirubin with the
sources ranges between 250 to 300 mg/day, of which 85% is derived Stability:at 2 C to 8 C up to the expiration date labeled on the photoelectric colorimetric method. J Biol Chem.1937:119:481-
from the heme moiety of the haemoglobin released from senescent bottles. Linearity 490.
erythrocytes that are destroyed in the reticuloendothelial system. Always keep bottles closed tightly and protect from light. The reaction is linear up to Bilirubin concentration of 25 mg/dl 3. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
The remaining 15 % is produced from erythrocytes destroyed in the (427.5 mmol/l) ed. Philadephia: WB saunders; 1995:268-273.r P. Neue
bone marrow and from catabolism of other heme containing proteins Specimen collection and preparation serumeisenbestimmung auf dem GSA II. Lab Med.1980;4:240-
such as cytochromes and myoglobin. Avoide exposure of the specimen to light. If plasma is used, only Interfering substances 244.
After it is produced in the peripheral tissues, bilirubin is transported heparin and oxalate plasma are suitable. Other anticoagulants Serum, plasma 4. Williams HL, Johnson DJ, Haut MJ. Simultaneous
to the liver in association with albumin. In the liver, bilirubin is should not be used. The average half-life of total bilirubin and direct spectrophotometry of Fe2+ and Cu2+ in serum denatured with
guanidine hydrochloride. Clin Chem.1977;23:237-240.
conjugated with glucuronic acid for solubilization and subsequent bilirubin in serum is 17 days and few hours respectively. Haemolysis
transport through the bile duct and elimination via the digestive Avoid haemolysis since it interferes with the test. 5. Makino et al.: Clinica Chimica Acta, 171 (1988), 19-28.
tract. Disease or conditions which, through hemolytic processes, Deterioration
produce bilirubin faster than the liver can metabolize it, cause the Do not use the Spectrum bilirubin reagents if precipitate forms. Lipemia
levels of unconjugated (indirect) bilirubin to increase in the circulation. Failure to recover control values within the assigned range may be Lipemic specimens interfere with the test.
Bile duct obstruction or damage to hepatocellular structure causes an indication of reagent deterioration. ORDERING INFORMATION
increases in the levels of both conjugated (direct) and unconjugated Drugs CATALOG NO QUANTITY
(indirect) bilirubin in the circulation. Materials Provided Theophyllin and propranolol may cause artificially low total bilirubin
221 001 2 x 50 ml
- Total Bilirubin Reagent levels. 221 002 2 x 100 ml
Method Additional Material may be required:
Modified Bergh & Muller method (Colorimetric, End point) - Bilirubin Calibrators Expected Values
This reagent can be used manually (see methods below) and on
Assay Principle most Autoanalyzers, Applications are available on request. Total Bilirubin
The azobilirubin produced by the reaction between bilirubins and the
diazonium salt of 3,5-dichlororphenyl diazonium tetrafluoroborate System Parameters Newborns premature (3-5 d) 10-14 mg/dL (171-239 mmol/l)
shows maximum absorption at 540 nm. The intensity of the colour Wavelength 546 nm
produced is proportional to the quantity of bilirubin which has Optical path 1 cm Newborns:
reacted.In the presence of caffeine and surfactants as accelerators Assay type Colorimetric End-point (3-5 d) 4.0 - 8.0 mg/dL ( 68-137 mmol/l)
conjugated and free bilirubin participate in the reaction in the same Direction Increase (<48 h) 6.0 - 10.0 mg/dL (103-171 mmol/l)
o o
way, so that the level of total bilirubin is determined. Temperature 25 C or 37 C (<24 h) 2.0-6.0 mg/dL (34-103 mmol/l)
Zero adjustment Reagent blank
Reagents Sensitivity 0.1 mg/dL (1.71 mmol/l)
Linearity 25 mg/dL (427.5 mmol/l) Spectrum Diagnostics does not interpret the results of a
Reagent (R) clinical laboratory procedure; interpretation of the reults is
3,5-dichlorophenyl diazonium tetrafluoroborate 0.2 mmol/L Procedures considered the responsibility of qualified medical personnel.
All indications of clinical significance are supported by
Caffeine 50 mmol/L Blank Serum Blank Sample literature references.
Surfactants and stabilizers < 3% Serum blank
------- 1 ml -------
reagent (S)
Serum Blank Reagent (SB) Reagent (R) 1 ml ------- 1 ml Analytical Range
Buffered Saline 0.1 - 25 mg/dl.
Sample ------- 25 ml 25 ml

o o
For further information, refer to the Pediatric Total Bilirubin reagent Mix well and let stand 5 minutes at 25 C or 2 min at 37 C , then
material safety data sheet. read absorbance of sample blank and sample against reagent blank.
14 15
TOTAL Bilirubin SYMBOLS IN PRODUCT LABELLING Procedure 2 ( with Bilirubin Calibrator ) Sensitivity
(Single Reagent) EC REP Authorised Representative
IVD For in-vitro diagnostic use
Temperature Limitation
Use by/Expiration Date
Blank Calibrator sample blank sample The sensitivity of the reagent is 0.1 mg/dl. The lower detection limit
represents the lowest measurable Bilirubin concentration that can
MODIFIED BERGH & MÜLLER METHOD LOT Batch Code/Lot number CAUTION. Consult instructions
Samlpe ------ ------ 50 ml 50 ml
be distinguished from zero.
REF Catalogue Number for use Reagent 1 ml 1 ml ------ 1 ml
Consult instructions for use Manufactured by Calibrator ------ 50 ml ------ ------
Linearity
REF: 223 001 ( 4 x 25 ml) 100 test N.Saline ------ ------ 1 ml ------ The reaction is linear up to a total bilirubin concentration of 25 mg/dl.
REF: 223 002 ( 2 x 100 ml) 200 test Specimens showing higher concentration should be diluted
o o
Mix and incubate for 5 minutes at 15 -25 C or 3 minutes At 37 C. 1+4 with physiological saline and repeat the assay (result × 5)
Intended Use Reagent Preparation, Storage and Stability Measure absorbance of sample (Asample) Calibrator (Acal)and Interfering Substances
Spectrum Diagnostics Total Bilirubin single reagent is intended Spectrum Total Bilirubin Reagent is supplied ready to use. Hemolysis: Elevated levels of haemoglobin may interfere. Lipemia
for the In-vitro quantitative, diagnostic determination of bilirubin in Stability: at +2°C to +8°C up to the expiration date Sample blank (Asample blank) against reagent blank. (Intralipid): Elevated levels of triglycerides may interfere.
humanSerum on Stat Lab Instrument.
Deterioration Calculation Expected Values
Background Do not use the Spectrum bilirubin reagents if precipitate forms . DA Sample = Asample - Asample blank Serum: 0.1 to 1.2 mg/dl .
The average level of the bilirubin produced in humans from different Failure to recover control values within the assigned range may be Each laboratory should investigate the transferability of the expected
(Asample)
sources range between 250 to 300 mg/day, of which 85 % is derived an indication of reagent deterioration . T.Bilirubin concentration (mg/dl) = × conc. of cal values to its own patient population and if necessary determine its
(Acal)
from the heme moiety of the haemoglobin released from senescent own reference range.
erythrocytes that are destroyed in the reticuloendothelial system . Specimen Collection and Preservation
The remaining 15 % is produced from erythrocytes destroyed in the Avoide exposure of the specimen to light. If plasma is used, only Bilirubin calibrator is not included in the kit. Any commercial Bilirubin Spectrum Diagnostics does not interpret the results of a
bone marrow and from catabolism of other heme containing proteins heparin and oxalate plasma are suitable. Other anticoagulants calibrator is required for the test ( procedur2) clinical laboratory procedure; interpretation of the results is
such as cytochromes and myoglobin . should not be used. The average half-life of total bilirubin and considered the responsibility of qualified medical personnel.
After it is produced in the peripheral tissues , bilirubin is transported direct bilirubin in serum is 17 days and few hours respectively . Important Note :
to the liver in association with albumin . In the liver, bilirubin is For severely haemolyzed or lipemic sera serum correction
conjugated with glucuronic acid for solubilization and subsequent System Parameters is required by performing serum blank. Use normal saline as
transport through the bile duct and elimination via the digestive tract. Wavelength 546 nm serum blank reagent. Dynamic Range
Disease or conditions which, through hemolytic processes, produce Optical path 1 cm 0.1 - 25 mg/dL .
bilirubin faster than the liver can metabolize it, cause the levels of Assay type End-point Quality Control
unconjugated (indirect) bilirubin to increase in the circulation. Bile Direction Increase Normal & abnormal commercial control serum of known Waste Disposal
duct obstruction or damage to hepatocellular structure causes Sample : Reagent Ratio 1 : 20 concentrations should be analyzed with each run. This product is made to be used in professional laboratories.
increases in the levels of both conjugated (direct) and unconjugated Please consult local regulations for a correct waste disposal.
(indirect) bilirubin in the Temperature
o
37 C or 20 – 25 C
o
Performance Characteristics S56: dispose of this material and its container at hazardous or
circulation. Incubation time
o
5 minutes at 20 – 25 C Precision special waste collection point.
Zero adjustment Reagent Blank S57: use appropriate container to avoid environmental
Method Reagent Blank Limits Low 0.00 AU Within run (Repeatiblity) contamination.
MODIFIED BERGH & MÜLLER METHOD (Colorimetric, End Point) High 0.15 AU Level 1 Level 2 S61: avoid release in environment. refer to special
Sensitivity 0.1 mg/dL n 20 20 instructions/safety data sheets.
Assay Principle Linearity 25 mg/dL
Mean (mg/dL) 0.79 4.37
The azobilirubin produced by the reaction between bilirubins and References
SD 0.016 0.18
the diazonium salt of 3,5 dichlorophenyl tetraflouroborate salt shows Procedure 1 ( with factor ) 1. Balistreri WF, Shaw LM. Liver function. In: Tietz NW, ed.
maximum absorption at 540 nm. The intensity of the colour produced CV% 2.13 4.12 Fundamentals of clinical chemistry.3 rd ed. Philadelphia:WB
is proportional to the quantity of bilirubin which has reacted. In the Blank sample blank sample Saunders: 1987:729-761.
presence of caffeine and surfactants as accelerators conjugated and Samlpe ------ 50 ml 50 ml Run to run (Reproducibility) 2. Malloy HT, Evelyn KA. The determination of bilirubin with the
free bilirubin participate in the reaction in the same way, so that the photoelectric colorimetric method. J Biol Chem.1937:119:481--
Reagent 1 ml ------ 1 ml 490.
level of total bilirubin is determined. Level 1 Level 2
N.Saline ------ 1 ml ------
n 20 20 3. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
Reagent (R) ed.Philadephia: WB saunders; 1995:268-273.
o o Mean (mg/dL) 0.79 4.37
3,5-dichlorophenyl tetrafluoroborate 0.2 mmol/l Mix and incubate for 5 minutes at 15 -25 C or 3 minutes At 37 C.
Measure absorbance of sample (Asample) and Sample blank SD 0.016 0.18
Caffeine 50 mmol/l
Surfactants and stabilizers < 3% (Asample blank) against reagent blank. CV% 2.13 4.12
ORDERING INFORMATION
CATALOG NO QUANTITY
For further information, refer to the Total Bilirubin reagent material Calculation Methods Comparison
223 001 100 Test
safety data sheet. 1- with factor A comparison of the Spectrum BIL-T (y) with a commercial obtainable 223 002 200 Test
DA Sample = Asample - Asample blank assay (x) gave the following result:
Precautions and Warnings Total Bilirubin Factor = 28 y = 0.999 x + 0.250; r = 0.997
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Serum Total Bilirubin Conc (mg/dl) = DA Sample x 28
immediately with plenty of soap and water. In case of severe injuries; Conversion Factor = mg/dl x 17.1 = µmol/l
16 17
Bilirubin (DIRECT) SYMBOLS IN PRODUCT LABELLING
Run to run (Reproducibility) References
Direct 1. Balistreri WF, Shaw LM. Liver function. In: Tietz NW, ed.
Colorimetric EC REP Authorised Representative
Use by/Expiration Date Level 1 Level 2 Fundamentals of clinical chemistry.3 rd ed. Philadelphia:WB
Saunders: 1987:729-761.
REF: 224 001 120 Test LOT Batch Code/Lot number CAUTION. Consult instructions n 20 20
REF Catalogue Number for use 2. Malloy HT, Evelyn KA. The determination of bilirubin with the
Mean (mg/dL) 0.32 0.82
R1 Direct Bilirubin 2 x 90 ml Consult instructions for use Manufactured by photoelectric colorimetric method. J Biol Chem.1937:119:481-
R2 Nitrite 1 x 7 ml SD 0.023 0.062 490.
CV% 5.57 8.1 3. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
Specimen Collection and Preservation ed.Philadephia: WB saunders; 1995:268-273.
Avoide exposure of the specimen to light. If plasma is used, only
Intended Use heparin and oxalate plasma are suitable. Other anticoagulants Methods Comparison
Spectrum Diagnostics bilirubin reagent is intended for the in-vitro should not be used. The average half-life of total bilirubin and direct A comparison between Spectrum Diagnostics Bilirubin and a
quantitative, diagnostic determination of bilirubin in human serum bilirubin in serum is 17 days and few hours respectively. commercial reagent of the same methodology was performed on 20
on both automated and manual systems. human sera. A correlation of 0.975 was obtained.
Stability:
Background o o o Sensitivity CATALOG NO QUANTITY
The average level of the bilirubin produced in humans from different -20 C 4–8 C 20 – 25 C
When run as recommended, the sensitivity of this assay is 0.1 mg/dL 224 001 120 test
sources ranges between 250 to 300 mg/day, of which 85% is derived Direct 6 months 7 days 2 days (1.7 mmol/L) for total and 0.04 mg/dL (0.68 mmol/L) for direct bilirubin.
from the heme moiety of the haemoglobin released from senescent
erythrocytes that are destroyed in the reticuloendothelial system. Linearity
The remaining 15 % is produced from erythrocytes destroyed in the Procedure The reaction is linear up to a direct bilirubin concentration of 18 mg/
bone marrow and from catabolism of other heme containing proteins dL(308 mmol/L). Specimens showing higher concentration should be
such as cytochromes and myoglobin. Direct Bilirubin diluted 1+4 with physiological saline and repeat the assay (result×5).
After it is produced in the peripheral tissues, bilirubin is transported
to the liver in association with albumin. In the liver, bilirubin is Sample blank Sample
conjugated with glucuronic acid for solubilization and subsequent Reagent 1 (D) 1.5 ml 1.5 ml Serum, plasma
transport through the bile duct and elimination via the digestive
tract. Disease or conditions which, through hemolytic processes, Reagent 2 ----- 50 ml
Haemolysis
produce bilirubin faster than the liver can metabolize it, cause the Sample 100 ml 100 ml Avoid haemolysis since it interferes with the test.
levels of unconjugated (indirect) bilirubin to increase in the circulation. o
Mix and incubate for 5 minutes at 20 – 25 C. Measure absorbance
Bile duct obstruction or damage to hepatocellular structure causes
increases in the levels of both conjugated (direct) and unconjugated of sample (Asample) against sample blank at 546 nm(530 - 580 Lipemia
nm) Lipemic specimens interfere with the test.
(indirect) bilirubin in the circulation.
Drugs
Method Calculation
Theophyllin and propranolol may cause artificially low total bilirubin
Colorimetric method. levels.
{(A)Sample - (A) Sample blank} x Factor* = mg/dl
Assay Principle Expected Values
Bilirubin is converted to colored diazotized sulfanilic acid and *Theoritical Factor
measured photometrically. Of the two fractions presents in serum, Direct Bilirubin 0 – 0.3 mg/dL (0 – 51 mmol/L)
bilirubin glucuromide and free bilirubin loosely bound to albumin. Direct bilirubin = 14
only the former reacts directly in aqueous solution (bilirubin direct),
Conversion Factor = mg/dl x 17.1 = mmol/l Spectrum Diagnostics does not interpret the results of a
clinical laboratory procedure; interpretation of the results is
Reagents
Note considered the responsibility of qualified medical personnel.
Reagent 1 (R1) D- Bilirubin
For bilirubin determination in newborns, pippete 50 ml of sample. literature references.
Sulfanilic acid 30 mmol/l
HCL 150 mmol/l Multiply the result by 2.
Analytical Range
Reagent 2 (R2)
Sodium Nitrite 29 mmol/l Quality Control
Direct bilirubin : 0.04 – 18 mg/dL ( 0.68 – 308 mmol/L)
Precautions and Warnings concentrations should be analyzed with each run
Waste Disposal
S28 After contact with skin, wash immediately with plenty of Performance Characteristics
This product is made to be used in professional laboratories.
soap and water. Precision Please consult local regulations for a correct waste disposal.
Within run (Repeatiblity)
Do not ingest or inhalate. In case of contact with eyes or skin; rinse special waste collection point.
immediately with plenty of soap and water. In case of severe injuries; Direct S57: use appropriate container to avoid environmental
seek medical advice immediately. contamination.
Level 1 Level 2
Reagent Preparation, Storage and Stability n 20 20 safety data sheets.
Spectrum bilirubin reagents are supplied ready-to-use and stable up
o Mean (mg/dL) 0.299 0.77
to the expiry date labeled on the bottles when stored at 2 - 8 C
SD 0.016 0.057
Deterioration CV% 5.41 7.4
Do not use the Spectrum bilirubin reagents if precipitate forms.
Failure to recover control values within the assigned range may be
an indication of reagent deterioration.
18 19
Bilirubin (TOTAL AND DIRECT) SYMBOLS IN PRODUCT LABELLING
Quality Control
Spectrum Diagnostics does not interpret the results of a
DMSO, Colorimetric EC REP Authorised Representative
concentrations should be analyzed with each run.Performance
Characteristics
considered the responsibility of qualified medical personnel.
LOT Batch Code/Lot number CAUTION. Consult instructions literature references.
REF: 225 001 60 Test REF: 225 002 150 Test REF Catalogue Number for use Precision
R1 Direct Bilirubin 1x90 ml R1 Direct Bilirubin 1x225 ml Within run (Repeatiblity) Analytical Range
R2 Total Bilirubin 1x90 ml R2 Total Bilirubin 1x225 ml Total bilirubin : 0.1 – 18 mg/dL ( 1.7 – 308 mmol/L)
Total Direct
R3 Nitrite 2 x 7 ml R3 Nitrite 2 x 10 ml Direct bilirubin : 0.04 – 18 mg/dL ( 0.68 – 308 mmol/L)
Reagent Preparation, Storage and Stability Level 1 Level 2 Level 1 Level 2
Spectrum bilirubin reagents are supplied ready-to-use and stable up
REF: 225 003 200 Test o n 20 20 20 20 Waste Disposal
to the expiry date labeled on the bottles when stored at 2 - 8 C
R1 Direct Bilirubin 3x100 ml Mean (mg/dL) 0.79 4.37 0.299 0.77 Please consult local regulations for a correct waste disposal.
R2 Total Bilirubin 3x100 ml Deterioration SD 0.016 0.18 0.016 0.057 S56: dispose of this material and its container at hazardous or
R3 Nitrite 1 x 20 ml Do not use the Spectrum bilirubin reagents if precipitate forms. special waste collection point.
Failure to recover control values within the assigned range may be CV% 2.13 4.12 5.41 7.4 S57: use appropriate container to avoid environmental
Intended Use an indication of reagent deterioration. contamination.
Spectrum Diagnostics bilirubin reagent is intended for the in-vitro S61: avoid release in environment. refer to special instructions/
Specimen Collection and Preservation Run to run (Reproducibility) safety data sheets.
quantitative, diagnostic determination of bilirubin in human serum
on both automated and manual systems. Avoide exposure of the specimen to light. If plasma is used, only Total Direct
heparin and oxalate plasma are suitable. Other anticoagulants References
should not be used. The average half-life of total bilirubin and direct Level 1 Level 2 Level 1 Level 2
Background 1. Balistreri WF, Shaw LM. Liver function. In: Tietz NW, ed.
The average level of the bilirubin produced in humans from different bilirubin in serum is 17 days and few hours respectively. n 20 20 20 20 Fundamentals of clinical chemistry.3 rd ed. Philadelphia:WB
sources ranges between 250 to 300 mg/day, of which 85% is derived Mean (mg/dL) 0.82 4.52 0.32 0.82 Saunders: 1987:729-761.
from the heme moiety of the haemoglobin released from senescent Stability: 2. Malloy HT, Evelyn KA. The determination of bilirubin with the
erythrocytes that are destroyed in the reticuloendothelial system. o o o SD 0.02 0.27 0.023 0.062 photoelectric colorimetric method. J Biol Chem.1937:119:481-
-20 C 4–8 C 20 – 25 C
The remaining 15 % is produced from erythrocytes destroyed in the CV% 2.24 4.21 5.57 8.1 490.
bone marrow and from catabolism of other heme containing proteins Total 6 months 7 days 1 day 3. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
such as cytochromes and myoglobin. Direct 6 months 7 days 2 days ed.Philadephia: WB saunders; 1995:268-273.
After it is produced in the peripheral tissues, bilirubin is transported Methods Comparison
to the liver in association with albumin. In the liver, bilirubin is Procedure A comparison between Spectrum Diagnostics Bilirubin and a
conjugated with glucuronic acid for solubilization and subsequent commercial reagent of the same methodology was performed on 20
transport through the bile duct and elimination via the digestive Direct Bilirubin human sera. A correlation of 0.975 was obtained. ORDERING INFORMATION
tract. Disease or conditions which, through hemolytic processes,
produce bilirubin faster than the liver can metabolize it, cause the Sample blank Sample CATALOG NO QUANTITY
Sensitivity
levels of unconjugated (indirect) bilirubin to increase in the circulation. Reagent 1 (D) 1.5 ml 1.5 ml When run as recommended, the sensitivity of this assay is 0.1 mg/dL 225 001 60 test
Bile duct obstruction or damage to hepatocellular structure causes (1.7 mmol/L) for total and 0.04 mg/dL (0.68 mmol/L) for direct bilirubin. 225 002 150 test
increases in the levels of both conjugated (direct) and unconjugated Reagent 3 ----- 50 ml
225 003 200 test
(indirect) bilirubin in the circulation. Sample 100 ml 100 ml Linearity
o
Mix and incubate for 5 minutes at 20 – 25 C. Measure absorbance The reaction is linear up to a total bilirubin concentration of 18 mg/
Method dL (308 mmol/L) and a direct bilirubin concentration of 18 mg/dL
of sample (Asample) against sample blank at 546 nm(530 - 580
DMSO. Colorimetric method. (308 mmol/L). Specimens showing higher concentration should be
nm)
diluted 1+4 with physiological saline and repeat the assay (result×5).
Assay Principle
Bilirubin is converted to colored diazotized sulfanilic acid and Interfering substances
measured photometrically. Of the two fractions presents in serum, Total Bilirubin
Serum, plasma
bilirubin glucuromide and free bilirubin loosely bound to albumin. Sample blank Sample
only the former reacts directly in aqueous solution (bilirubin direct),
Reagent 2 (T) 1.5 ml 1.5 ml Haemolysis
while free bilirubin requires solubilization with dimethylsulfoxide
Avoid haemolysis since it interferes with the test.
(DMSO) to react (bilirubin indirect). In the determination of indirect Reagent 3 ----- 50 ml
bilirubin the direct is also determined, the results correspond to total Sample 100 ml 100 ml Lipemia
bilirubin.
Mix and incubate for exactly 5 minutes at 20 – 25 C. Measure
o Lipemic specimens interfere with the test.
Reagents absorbance of sample (Asample) against sample blank at 546 nm
Drugs
(530 - 580 nm). Theophyllin and propranolol may cause artificially low total bilirubin
Reagent 1 (R1) D- Bilirubin levels.
Sulfanilic acid 30 mmol/l
HCL 150 mmol/l Calculation Expected Values
{(A)Sample - (A) Sample blank} x Factor* = mg/dl
Reagent 2 (R2) T- Bilirubin
Total Bilirubin
Sulfanilic acid 30 mmol/l *Theoritical Factor
HCL 150 mmol/l
Adults and infants>1 month < 0.2-1.1 mg/dL ( 3.4-17 mmol/l)
Dimethylsulfoxide(DMSO) 7 mol/l Direct bilirubin = 14 Newborns premature (3-5 d) 10-14 mg/dL (171-239 mmol/l)
Total bilirubin = 19.1
Reagent 3 (R3)
Newborns:
Sodium Nitrite 29 mmol/l Conversion Factor = mg/dl x 17.1 = mmol/l (3-5 d) 4.0 - 8.0 mg/dL ( 68-137 mmol/l)
(<48 h) 6.0 - 10.0 mg/dL (103-171 mmol/l)
Note (<24 h) 2.0-6.0 mg/dL (34-103 mmol/l)
Precautions and Warnings For bilirubin determination in newborns, pippete 50 ml of sample.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Multiply the result by 2. Direct Bilirubin 0 – 0.3 mg/dL (0 – 51 mmol/L)
immediately with plenty of soap and water. In case of severe injuries;
20 21
Cholesterol – Liquizyme SYMBOLS IN PRODUCT LABELLING
Procedure Drugs
Of the drugs tested in vitro, methyldopa causes artificially Low total
Blank Standard Sample
CHOD-PAP (Single Reagent) EC REP Authorised Representative
Use by/Expiration Date Reagent (R) 1.0 ml 1.0 ml 1.0 ml
cholesterol values at the tested drug Level.
Batch Code/Lot number CAUTION. Consult instructions Others

LOT Standard ------- 10 ml -------
Catalogue Number for use
Physiological ascorbic acid concentration does not interfere with
REF: 230 001 ( 2 x 25 ml) 50 test REF
Sample ------- ------- 10 ml the test. Ascorbic Acid levels higher than 425 mmol/l (7.5 mg/dl)
REF: 230 002 ( 4 x 25 ml) 100 test Consult instructions for use Manufactured by
decrease the apparent total cholesterol concentration significantly.
REF: 230 003 ( 4 x 30 ml) 120 test o
Mix and incubate for 5 minutes at 37 C or 10 minutes at
REF: 230 004 (10 x 15 ml) 150 test Reagent (R) 15 – 25 C. Measure absorbance of specimen (Aspecimen) and
o
Expected Values
REF: 230 005 ( 4 x 50 ml) 200 test standard (Astandard) against reagent blank within 30 minutes.
REF: 230 006 ( 4 x 100 ml) 400 test Pipes Buffer pH 7.0 50 mmol/L
The following guidelines may be used for clinical interpretation:
REF: 230 007 ( 8 x 100 ml) 800 test Phenol 30 mmol/L Calculation
REF: 230 008 ( 6 x 100 ml) 600 test (Aspecimen) Risk classification Total cholesterol
Sodium cholate 5.0 mmol/L Serum cholesterol conc. (mg/dL)= x 200
REF: 230 009 ( 2 x 500 ml) 1000 test (Astandard) Desirable <200 mg/dl <5.2 mmol/L
Cholesterol esterase >250 U/L
Quality Control Borderline high 200-239 mg/dl 5.2-6.2 mmol/L
Cholesterol oxidase >500 U/L Normal & abnormal commercial control serum of known
concentrations should be analyzed with each run. High ≥240 mg/dl ≥6.2 mmol/L
Peroxidase >2.0 KU/L
Intended Use 4-amino-antipyrine 1.0 mmol/L
Spectrum Diagnostics liquizyme cholesterol reagent is intended
Sodium Azide 8.0 mmol/L Precision Spectrum Diagnostics does not interpret the results of a
for in-vitro quantitative, diagnostic determination of cholesterol in
clinical laboratory procedure; interpretation of the reults is
human serum on both manual and automated systems.
For further information, refer to the Cholesterol reagent material Within run (Repeatiblity) considered the responsibility of qualified medical personnel.
safety data sheet. All indications of clinical significance are supported by
Background Level 1 Level 2 literature references.
Measurement of serum cholesterol levels is important as an indicator
of liver function, intestinal absorption, biliary function and in the Precautions and Warnings n 20 20
diagnosis and classification of hyperlipoproteinemias. Do not ingest or inhalate. In case of contact with eyes or skin; rinse Mean (mg/dL) 149.8 252
immediately with plenty of soap and water. In case of severe injuries; Dynamic Range
Elevated cholesterol levels may occur with hypothyroidism,diabetes 5 - 750 mg/dl (0.13 - 19.5 mmol/L).
seek medical advice immediately. SD 1.69 1.91
and nephrotic syndrome. Elevated serum cholesterol levels correlate
well with the incidence of coronary artery diseases. Stress, age, Reagent (R) contains sodium azide which may react with copper or CV% 1.13 0.76
lead plumbing. Waste Disposal
gender, hormonal balance and pregnancy affect normal cholesterol
levels. Depressed levels are associated with hyperthyroidism and
Reagent Preparation, Storage and Stability Run to run (Reproducibility) Please consult local regulations for a correct waste disposal.
severe liver diseases.
Spectrum cholesterol reagents are supplied ready-to-use and stable S56: dispose of this material and its container at hazardous or
up to the expiry date labeled on the bottles. Once opened, the Level 1 Level 2 special waste collection point.
Method o S57: use appropriate container to avoid environmental
CHOD-PAP-enzymatic colorimetric method. opened vial is stable for 3 months at 2 - 8 C. n 20 20
contamination.
Mean (mg/dL) 157 259 S61: avoid release in environment. refer to special instructions/
Assay Principle Deterioration
The reagent is normally clear or pale pink. Do not use liquizyme SD 1.77 2.12 safety data sheets.
The series of the reactions involved in the assay system is as
follows: cholesterol reagent if it is turbid or if the absorbance is greater than CV% 1.23 0.97
0.15 at 546 nm. References
1. Cholesterol esters are enzymatically hydrolyzed by cholesterol 1. Ellefson RD and Caraway WT : Fundamentals of clinical
esterase (CE) to cholesterol and free fatty acids. Methods Comparison chemistry. Ed Tietz NW 1976; p506.
Specimen Collection and Preservation A comparison between Spectrum Diagnostics Cholesterol reagent 2. Flegg HM : Ann Clin Biochem 1963 ; 10 : 79.
Cholesterol CE Cholesterol It is recommended that prior to sample collection, patients should and a commercial reagent of the same methodology was performed
+ 3. NCEP expert panel, Arch Intern Med 1988; 148: 36–69
Esters be following their usual diet and be in their usual state of health. on 20 human sera. A correlation of 0.988 was obtained.
Fatty acids 4. Richmond. N., Clin. Chem. 1973; 19: 1350-1356.
Patients who are actually ill, losing weight, pregnant or have had
5. Roeschlau,P., Bernt. E. and Gruber. W.J., Clin. Chem Clin.
2. Free cholesterol, including that originally present, is then a myocardial infarction in the previous 3 months should be resch- Sensitivity Biochem. 1974; 12:403.
oxidized by cholesterol oxidase (CO) to cholest-4-en-3-one and eduled. Both fasting and non fasting samples can be used. Non When run as recommended, the minimum detection limit of the
o 6. Trinder, P, Ann. Clin. Biochem. 1969; 6: 24.
hydrogen peroxide. haemolysed serum or plasma can be stored at 4 C up to 7 days assay is 5 mg/dL (0.13 mmol/L).
o o 7. Young DS .et al. Clin Chem. 1975 ; 21.
prior to analysis, 5-7 days at 20-25 C, stable for 3 months at -20 C,
o
Cholesterol Cholest-4-en-3-one and at -70 C for several months. The only acceptable anticoaglulant
CHOD Linearity
+ + is heparin.
H2O2 The reaction is linear up to a cholesterol concentration of 750 mg/
O2
dl; specimens showing higher concentration should be diluted 1+1
System Parameters ORDERING INFORMATION
using physiological saline and repeat the assay (result × 2).
3. The hydrogen peroxide combines with phenol and Wavelength 546 nm (500 – 550 nm) CATALOG NO QUANTITY
4-aminoantipyrine (4AAP) in the presence of peroxidase Optical path 1 cm
Interfering Substances: 230 001 2 x 25 ml
(POD) to form a chromophore (quinoneimine dye) which may Assay type End-point
be quantitated at 500 – 550 nm. For bichromatic analyzers the Direction Increase 230 002 4 x 25 ml
Haemolysis 230 003 4 x 30 ml
blank wavelength should be set to 600 or 650 nm. Sample : Reagent Ratio 1 : 100
No significant interference up to a level of 500 mg/dl. 230 004 10 x 15 ml
e.g .: Reagent volume 1 ml
Quinoneimine Dye Sample volume 10 ml 230 005 4 x 50 ml
2H2O2 +Phenol POD Icterus
+ + Temperature
o o
15 – 25 C or 37 C 230 006 4 x 100 ml
(4AAP) 4H2O No interference from free bilirubin up to a level of 15 mg /dL 230 007 8 x 100 ml
Zero adjustment Reagent blank
o (260 mmol/L) and conjugated bilirubin up to a level of 7 mg/dL 230 008 6 x 100 ml
Incubation time 5 minutes at 37 C or
o (116 mmol/L). 230 009 2 x 500 ml
Reagents 10 minutes at 15 – 25 C
Reagent Blank Limits Low 0.00 AU
Lipemia
Standard cholesterol (ST) High 0.15 AU
No significant interference up to 1.7 AU.
200 mg/dL 5.17 mmol/L Sensitivity 5 mg/dL (0.13mmol/L)
Linearity 750 mg/dL (19.5 mmol/L)
22 23
HDL CHOLESTEROL - SYMBOLS IN PRODUCT LABELLING
Clinical Interpretation
Standard Increased
Precipitant EC REP Authorised Representative
Desirable
Risk Level Risk Level

HDL Cholesterol
Females (mg/dL) >65 45 - 65 <45
REF: 266 001 (1 X 50 ml) 100 test REF Catalogue Number for use
(mmol/L) >1.68 1.16 - 1.68 <1.16
REF: 266 002 (4 X 100 ml) 800 test Consult instructions for use Manufactured by
Males (mg/dL) >55 35 - 55 <35
REF: 266 003 (2 X 50 ml) 200 test (mmol/L) >1.42 0.90 - 1.42 <0.90
REF: 266 004 (5 X 50 ml) 500 test Specimen Collection and Preservation LDL Cholesterol
(mg/dL) <150 150 - 190 >190
Serum or plasma (mmol/L) <3.38 3.88 - 4.91 >4.91
Intended Use EDTA and Heparin may be used as anticoagulants.
Spectrum Diagnostics HDL cholesterol reagent is intended for Total Cholesterol
the in-vitro quantitative, diagnostic determination of HDL cholesterol o (mg/dL) <200 200 - 300 >300
Stability : 7 days at 2 – 8 C
in human serum , heparinized or EDTA plasma . o (mmol/L) <5.17 5.17 - 7.76 >7.76
4 days at 20 - 25 C
Background Procedure
High density lipoprotein measurement, in conjunction with other lipid Spectrum Diagnostics does not interpret the results of a
1 - Precipitation
determination, has been shown to be useful in assessing the risk of clinical laboratory procedure ; interpretation of the results is
coronary heart disease. HDL is responsible for carrying cholesterol considered the responsibility of qualified medical personnel.
Pipette into centrifuge tubes :
back from preipheral cells to the liver , therefore the risk of coronary All indications of clinical significance are supported by
heart disease is lowered with increased levels of HDL. usually, very Reagent 0.5 ml literature references .
low density lipoprotein (VLDL) and low density lipoprotein (LDL) are Specimen 0.2 ml
selectivety precipitated from serum or plasma samples followed by
determination of cholesterol in the HDL-containing supernatant. Mix and incubate for 10 minutes at room temperature, then centrifuge References
for 10 minutes at 4000 rpm .
Method Carefully collect the supernatant . 1. National Cholesterol Education Program Recommendation
Precipitation Method . o
Stability : the supernatant may be stored up to five days at 2 - 8 C for Measurement of High-Density Lipoprotein Cholesterol:
Executive Summary. Clin Chem. 1995;41:1427 - 1433.
Assay Principle 2 - Cholesterol - Liquizyme
low density lipoproteins (LDL) and very low density lipoproteins 2. Friedewald , W.T. et al. Clin. Chem. 1972; 18: 499.
(VLDL) in sample precipitate with phosphotungstate and magnesium Pipette into test tubes : 3. Lopes- Virella, M.F. et al. Clin. Chem. 1977; 23: 882.
ions. After centrifugation, the cholesterol concentration in the HDL
fraction, which remains in the supernatant, is determined. Blank Specimen
Distilled water 50 ml -------
Cholesterol esters + H2O chol. esterase Cholesterol + Fatty acid ORDERING INFORMATION
Specimen supernatant ------- 50 ml
REAGENTS
Cholesterol + 1\2 O2 + H2O chol. oxidase Cholestenone + H2O2 Cholesterol Reagent 1ml 1ml
CATALOG NO QUANTITY
peroxidase
2H2O + 4-Aminoantipyrine + phenol Quinoneimine + 4H2O o
Mix, incubte for 10 minutes at 20 – 25 C or 5 minutes at 37 C.
o
266 001 1 x 50 ml
Measure the absorbance of the specimen (Aspecimen) against 266 002 4 x 100 ml
Reagents reagent blank at 546 nm (500 – 550 nm) within 60 minutes. 266 003 2 x 50 ml
266 004 5 x 50 ml
Reagent (R)
Phosphotungstate 0.52 mmol/L Calculation
Magnesium chloride 30 mmol/L HDL cholesterol conc. (mg/dL) = Asample x 570
Reagents also contain non-reactive stabilizers and Expected Values

surfactants . Females 48.6 - 75 mg/dL 1.26 - 1.94 mmol/L
Males 41.0 -58.7 mg/dL 1.06 - 1.52 mmol/L
* Supplementary reagents : Apack for Spectrum liquizyme Childern 51.8 - 71.9 mg/dL 1.34 - 1.86 mmol/L
cholesterol reagent is required
To calculate LDL cholesterol
Pay attention to all precautions and warnings listed in Spectrum in mg/dL
Diagnostics
catalogue available upon request . LDL Total Triglycerides HDL
Cholesterol = Cholesterol - 5 - Cholesterol
Reagent Preparation, Storage and Stability
Spectrum HDL cholesterol reagent is supplied ready-to-use
and stable up to the expiry date labeled on the bottles when in mmol/L
o
properly stored at 2 - 8 C.
LDL Total Triglycerides HDL
Deterioration Cholesterol = Cholesterol - 2.2 - Cholesterol
Do not use The HDL cholesterol reagents if precipitate forms .
Failure to
recover control values within the assigned range may be an
indication of reagent deterioration .
24 25
HDL CHOLESTEROL SYMBOLS IN PRODUCT LABELLING
QUALITY CONTROL
Control sera are recommended to monitor the performance of assay
Direct Enzymatic colorimetric, Liquid EC REP Authorised Representative Temperature Limitation procedures. CATALOG NO QUANTITY
IVD For in-vitro diagnostic use Use by/Expiration Date If control values are found outside the defined range, check the 267 001 100 tests
REF: 267 001 100 Test REF: 267 002 200 Test LOT Batch Code/Lot number CAUTION. Consult instructions instrument reagents and calibrator for problems. 267 002 200 tests
R1 1 x 30 ml R2 1 x 10 ml R1 2 x 30 ml R2 1 x 20 ml REF Catalogue Number for use Each laboratory should establish its own Quality Control scheme and
corrective actions if controls do not meet the acceptable tolerances.
REAGENT PREPARATION AND STORAGE Expected Values Men Women

Intended use
• R1 and R2: Are ready to use. Low risk > 50 mg/dL > 60 mg/dL
Spectrum diagnostics HDL cholesterol reagent is intended for in-
vitro quantitative determination of HDL cholesterol in human serum, • HDL CAL: Dissolve the contents with distilled water, as Normal risk 35-50 mg/ dL 45-60 mg/dL
heparinized or EDTA plasma. mentioned on vial label. Cap vial and mix gently to dissolve High risk < 35 mg/dL < 45 mg/dL
contents. Wait for 30 minutes before use.
Background o These values are for orientation purpose; each laboratory should
HDL particles serve to transport in the blood-stream. • R1 and R2: Once opened is stable 8 weeks at 2-8 C.
establish its own reference range.
HDL is known as “good cholesterol” because high levels are thought • HDL CAL: Once reconstituted 1 week at 2-8 C.
o
to lower the risk of heart disease and coronary artery disease. A

low HDL cholesterol level, is considered a greater heart disease
o
or 4 weeks at -20 C. Dynamic range:
The measuring range is from 1.0 mg/dL to linearity
risk.1.5.6. • Do not use reagents over the expiration date. limit of 180 mg/dL. If the results obtained were greater than linearity
Clinical diagnosis should not be made on a single test result; it
limit, dilute the sample 2 times with NaCl 9 g/L and multiply the result
Should integrate clinical and other laboratory data. • Sign of reagent deterioration.
by 2.
• Presence of particles and turbidity.
METHOD
Direct Enzymatic colorimetric
Sensitivity
The sensitivity of the test is 1 mg/dl
ADDITIONAL EQUIPMENT
PRINCIPLE • Spectrophotometer or colorimeter measuring at 600 nm.
The assay is based on a modified polyvinyl sulfonic acid (PVS)
Accuracy
• Matched cuvettes 1.0 cm light path. Results obtained using Spectrum reagents (y) did not show
and polyethylene-glycol-methyl ether (PEGME) coupled classic
systematic difference when compared with other commercial
precipitation method with the improvements in using optimized • General laboratory equipment.
reagents. (x).
quantities of PVS/PEGME and selected detergents.LDL, VLDL,
The results obtained using 50 samples were the following. Correction
and chylomicron (CM) react with PVS and PEGME and the reaction
REAGENT STABILITY coefficient (r): 0.996.
results in inaccessibility of LDL, VLDL and CM by cholesterol
All the components of the kit are stable until the expiration date on Regression equation: y 0.98 + 3.42 mg/dL.
Oxidase (CHOD) and cholesterol esterase (CHER). The enzymes
selectively react with HDL to produce H2O2 which is detected through the label when stored tightly closed at 2-8°C and contaminations are
a Tinder reaction. prevented during their use. Do not freeze the reagents. INTERFERENCES
No Interferences were observed to bilirubin T. and D. up to 60 mg/dL.
HDL + LDL + VLDL + CM PVS
HDL + (LDL + VLDL + CM) LINEARITY: 150 mg/dl Hemoglobin up to 1000 mg/dL or lipaemia up to 1800 mg/dL.
PEGME
HDL + CHOD + CHER PVS/PEGME

Fatty Acid + H2O2 PROCEDURE NOTES
This reagent can be used manually (see method below) and on Spectrum has Instrument application sheets for several automatic
Peroxidase analyzers. Instructions for many of them are available on request.
H2O2 +4-AA+TODB Quenone + 5 H2O most analyzers. Applications are available on request.
(λmax=560nm)
Wavelength 600 nm ( 580 nm is an option) Spectrum Diagnostics does not interpret the results of a
REAGENTS Cuvette 1 cm clinical laboratory procedure ; interpretation of the results is
Temperature 37 °C considered the responsibility of qualified medical personnel.
Reagents Composition Measure Against distelled water All indications of clinical significance are supported by
literature references .
Reagent 1 (R1): MES buffer (pH 6.5), TODB N, N-Bis (4-
Blank Calibrator Sample
sulfobutyl)-3- methylaniline), Polyvinyl sulfonic acid, Polyethylene-
glycol-methyl ester, MgCl2, Detergent, EDTA R1 (μL) 300 300 300 BIBLIOGRAPHY
Calibrator(μL) ------- 4 ------- 1. Natio H KCholesterol Kaplan A et al. Clin Chem the C.V. Mosby
Reagent 2(R2): MES buffer (pH 6.5), Cholesterol esterase, Co. St Louis. Toronto. Princeeton 1984; 1207-1213 and 437.
Cholesterol Oxidase, Peroxidase, 4-aminoantipyrine, detergent. Sample (μL) ------- ------- 4 2. US National Cholestrol Educatiopn Program of the National
Mix and Incubate for 5 min at 37°C. Then add : Institutes of Health.
HDL CAL 3. Young DS. Effects of Drugs on Clinical Lab. Tests, 4th ad AACC
Standard, Lyophilized Human Serum R2(μL) 100 100 100 Press, 1995.
4. Young DS. Effects of diseases on Clinical Lab. Tests 4th ad
Precautions Mix Read immediately the absorbance (A1) of the samples and AACC 2001.
HDL. CAL calibrator, against the Blank, then Read the absorbance (A2) of the 5. Burlis A et al. Tietz Texbook of Clinical Chemistry, 3rd ed AACC
Components from human origin have been tested and found to be Samples and calibrator after 5 mins, against the Blank. 1999.
negative for the presence of HBsAg, HCV and antibody to HIV (1/2). Calculate the Increase of the absorbance D A = A2 - A1. 6. Tietz N W et al, Clinical to Laboratory Tests, 3rd ed AACC 1995.
However handle cautiously as potentially infectious.
CALCULATION
SAMPLE COLLECTION AND PRESERVATION
(D A) Sample
Serum or heparinized plasma, free of haemolysis; Anticoagulants ────────── X Calibrator conc. = mg/dL of HDL-C
(D A) Calibrator
containing citrate should not be used.
Removed from the blood clot as soon as possible. Stability of the
o
sample: 7 days at 2-8 C. Conversion factor: mg/dL x 0.0259 = mmol/L
26 27
LDL CHOLESTEROL SYMBOLS IN PRODUCT LABELLING CALCULATION Spectrum Diagnostics does not interpret the results of a
EC REP Authorised Representative Temperature Limitation clinical laboratory procedure ; interpretation of the results is
DA Sample
Direct Enzymatic colorimetric, Liquid IVD For in-vitro diagnostic use Use by/Expiration Date X Calibrator conc. = mg/dL of LDL-C in the sample considered the responsibility of qualified medical personnel.
LOT Batch Code/Lot number CAUTION. Consult instructions DA Calibrator All indications of clinical significance are supported by
REF Catalogue Number for use literature references .
REF: 280 001 100 Test REF: 280 002 200 Test
R1 1 x 30 ml R2 1 x 10 ml R1 2 x 30 ml R2 1 x 20 ml QUALITY CONTROL
Control sera are recommended to monitor the performance of assay
BIBLIOGRAPHY
procedures.
1. Kaplan A et al. Lipoprotein Clin Chem the C. V. Masby Co. St
Intended Use Reagent Stability and Storage If control values are found outside the defined range, check the
Louis.
The SPECTRUM LDL-Cholesterol Assay is intended for the invitro Unopened reagents are stable until the expiration date printed on instrument reagents and calibrator for problems.
quantitative determination of Low Density Lipoprotein Cholesterol the outer box when stored at 2–8° C. Reagent on-board stability is Each laboratory should establish its own Quality Control scheme and 2. Okada M. et al Low- density lipoprotein can be chemically
measured J. LAb, Clin. Mad., 1996; 132, 195-201.
in human serum or plasma. The reagents can assist in the diagnosis at least 60 days. The reagent solutions should be clear. If turbid, the corrective actions if controls do not meet the acceptable tolerances.
and treatment of patients at risk of developing coronary reagents may have deteriorated. 3. Young DS. Effects of Drugs on Clinical Lab. Tests, 4th ad AACC
Press, 1995.
heart disease. Elevated LDL cholesterol is the primary target of Expected Values
cholesterol-lowering therapy. Specimen Collection and Preparation 4. Young DS. Effects of diseases on Clinical Lab. Tests 4th ad
AACC 2001.
Use fresh patient serum or plasma samples (EDTA, Citrate). If Levels of the risk
Clinical Significance samples contain LDL cholesterol greater than 250 mg/dL, they 5. Burlis A et al. Teitz Texbook of Clinical Chemistry, 3rd ed AACC
Low Density Lipoproteins (LDL) are synthesized in the liver by the should be diluted with saline. Desirable <100 mg/dL 1999.
action of various lipolytic enzymes on triglyceride-rich Very Low Medium 100 – 160 mg/dl 6. Tietz N W et al, Clinical Guide to Laboratory Tests, 3rd ed
Density Lipoproteins (VLDLs). Specific LDL receptors exist to LDL CAL: Dissolve the contents with the amount of distilled water High >160mg/dL AACC 1995.
facilitate the elimination of LDL from plasma by liver parenchymal indicated on Label. Cap vial and mix gently to dissolve contents, and
cells. It has been shown that most of the cholesterol stored in wait for 30 minutes. Stability is 7 days These values are for orientation purpose; each laboratory should
o o
atherosclerotic plaques originates from LDL. For this reason the At 2-8 C and 4 weeks at - 20 C. Establish its own reference range. ORDERING INFORMATION
LDL-Cholesterol concentration is considered to be the most CATALOG NO QUANTITY
important clinical predictor, of all single parameters, with respect to REAGENT STABILITY PERFORMANCE CHARACTERISTICS
280 001 100 tests
Coronary atherosclerosis. All the components of the kit are stable until the expiration 280 002 200 tests
date on the label when stored tightly closed at 2-8°C and Dynamic range:
Accurate measurement of LDL-Cholesterol is of vital importance contaminations are prevented during their use. Do not freeze the The measuring range is from 1.0 mg/dl to linearity
in therapies which focus on lipid reduction to prevent reagents. Limit of 250 mg/dl. If the results obtained were greater than linearity
atherosclerosis or reduce its progress and to avoid plaque rupture. limit, dilute the sample 2 times with NaCl 9 g/L and multiply the result
LINEARITY by 2.
Assay Principle 250 mg/dl
The assay is based on a modified polyvinyl sulfonic acid (PVS) Sensitivity:
and polyethylene-glycol methyl ether (PEGME) coupled classic PROCEDURE The sensitivity of the test is 1 mg/dl.
precipitation method with the improvements in using optimized This reagent can be used manually (see method below) and on
quantities of PVS/PEGME and selected detergents. LDL, VLDL, most analyzers. Applications are available on request. Accuracy: Results obtained using Spectrum reagents (y) did not
and chylomicron (CM) react with PVS and PEGME and the reaction show systematic differences when compared with other commercial
results in inaccessibility of LDL, VLDL and CM by cholesterol Wavelength 600 nm (580 nm is an option) reagents. (x).
oxidase Cuvette 1 cm
(CHOD) and cholesterol esterase (CHER), whereas HDL reacts Temperature 37 °C The results obtained using 92 samples were the following. Correction
with the enzymes. Addition of R2 containing a specific detergent Measure Against distelled water coefficient (r): 0.996.
releases LDL from the PVS/PEGME complex. The released LDL Regression equation: y =4.6+0.940x.
reacts with the enzymes to produce H2O2 which is quantified by the Blank Calibrator Sample
Tinder reaction. R1 (μL) 300 300 300 The results of the performance characteristics depend on the
analyzer used.
Calibrator(μL) ------- 4 -------
Reagent Composition
Sample (μL) ------- ------- 4
Reagent 1 (R1): MES buffer (pH 6.5) Polyvinylsulfonic INTERFERENCES
Acid Polyethyleneglycolmethylester, MgCl2 Detergent, EDTA, Mix and Incubate for 5 min at 37°C. Then add : No Interferences were observed with ascorbic acid up to 50 mg/dL,
4aminoantipyrine, Cholesterol esterase, Cholesterol oxidase R2(μL) 100 100 100 haemoglobin up to 500 mg/dL or bilirubin up to 30 mg/dL.
Peroxidase. A list of drugs and other interfering substances with LDL cholesterol
Mix Read immediately the absorbance (A1) of the samples and determination has been reported by Young et al 8.4
Reagent 2 (R2): MES buffer (pH 6.5), EDTA, Detergent calibrator, against the Blank, then Read the absorbance (A2) of the
TODB, N, N-Bis (4-sulfobutyl)-3-methylaniline) Samples and calibrator after 5 mins, against the Blank. NOTES
Calculate the Increase of the absorbance DA = A2 - A1. Spectrum has Instrument application sheets for several automatic
Reagent Preparation analyzers. Instructions for many of them are available on request.
SPECTRUM LDL-Cholesterol Assay Reagents (R1, R2) are liquid
stable, ready to use reagents.
28 29
Creatinine – Jaffè SYMBOLS IN PRODUCT LABELLING
Calculation
A2 – A1 = Aspecimen or Astandard.
Interfering Substances
Serum, plasma
EC REP Authorised Representative Use by/Expiration Date Concentration of creatinine in serum:
REF: 234 001 (2 x 100 ml) 200 test
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions (Aspecimen) Haemolysis
REF: 234 002 (4 x 100 ml) 400 test
LOT Batch Code/Lot number for use Creatinine (mg/dL) = x2 Erythrocyte contamination doesn’t elevate results.
REF: 234 003 (8 x 100 ml) 800 test (Astandard)
REF: 234 004 (2 x 500 ml) 1000 test REF Catalogue Number Manufactured by
Consult instructions for use (Xi) - Irritant
Icterus
REF: 234 005 (2 x 250 ml) 500 test
Concentration of creatinine in urine: Serum bilirubin levels higher than 5 mg/dL (85 mmol/L) decrease
REF: 234 006 (4 x 250 ml) 1000 test Temperature Limitation
(Aspecimen) serum creatinine.
Creatinine (mg/dL) = x 2 x 50
Intended Use (Astandard) Lipemia
Reagent Preparation
Spectrum Diagnostics creatinine reagent is intended for the in-vitro Lipemic specimens may cause high absorbance flagging. Diluted
Prepare working solution as following:
quantitative diagnostic determination of creatinine in human serum sample treatment may be recommended.
Combine one volume of R1 with one volume of R2 e.g. Creatinine clearance (ml/minutes):
or urine on both automated and manual systems.
1.0 ml R1 + 1.0 ml R2. mg creatinine / dl urine x ml urine / 24 hours
Expected Values
Background Reagent Storage and Stability mg creatinine / dl serum x 1440
Creatine is synthesized in kidney, liver and pancreas. It is All reagents are stable until expiration date stated on label when Serum, plasma
transported in blood to other organs such as muscle and brain where o Correction for body surface area can be done using the following Females 0.7-1.3 mg/dL 62-115 mmol/L
stored at 15 - 25 C. Working solution is stable for one day at
it is phosphorylated to phosphocreatine. Some free creatine in o formula for creatinine clearance: Males 0.9-1.5 mg/dL 80-133 mmol/L
15 – 25 C away from light.
muscle is converted to creatinine daily and the amount of creatinine Urine(24 hrs)
produced is proportional to muscle mass. In the absence of renal Serum creatinine / min. per standard surface area = Females 0.9 – 1.6 g/24 hrs
Deterioration
disease, excretion rate of creatinine in an individual is relatively UCr x V 1.73 Males 1.1 – 2.8 g/24 hrs
The creatinine reagents are not suitable for use if combined reagents x
constant. Therefore, measurement of creatinine clearance is useful
have an absorbance greater than 0.8 at 492 nm measured in a 1cm PCr A
in detecting renal disease and estimating the extent of impairment Creatinine clearance
lightpath or if the reagents develop a hazy appearance.
of renal function.Both serum creatinine and urea levels are elevated Females 75 – 115 ml / min.
in patients with renal malfunction, especially decreased glomerular Where: UCr = Concentration of creatinine in urine( mg/dl) Males 85 – 125 ml / min.
filtration. In the early stage of kidney damage, increase in serum urea Specimen Collection and Preservation PCr = Concentration of creatinine in plasma(mg/dl)
level usually precedes the increase in serum creatinine. However V = Volume of urine flow in mL/min.
Serum or plasma Spectrum Diagnostics does not interpret the results of a
serum urea levels may be affected by dehydration, diet and protein A = Body surface area in square meter .
Both are suitable for analysis. The only acceptable anticoagulants clinical laboratory procedure; interpretation of the results is
metabolism.On the other hand serum creatinine levels tend to be 1.73/A = Factor normalizes clearance for average body
are heparin and EDTA. Specimen should be promptly separated considered the responsibility of qualified medical personnel.
constant and unaffected by such factors. Thus serum creatinine is a surface.
from cells after blood collection. The biological half-life of creatinine All indications of clinical significance are supported by
significantly more reliable renal function screening test than serum
in blood is few minutes. literature references.
urea. o o
Note: Body surface area can be determined from height weight via
Stability: 7 day 2 - 8 C ; > 1 year at -20 C.
normograms in Tietz(6).
Method Analytical Range
Buffered Kinetic jaffé reaction without deproteinization. Urine
Thymol or toluene may be used for urine preservation. To determine
Quality Control 0.31 – 20 mg/dL (0.027-1.77 mmol/L).
Assay Principle creatinine concentration in urine, dilute 1 part sample with 49 parts
isotonic saline prior to assay. Multiply result by 50 to compensate
concentrations should be analyzed with each run. Waste Disposal
Creatinine reacts with picric acid under alkaline condition to form This product is made to be used in professional laboratories.
a yellow-red complex. The absorbance of the color produced, for dilution.
o
Stability: 2 days at 15 - 25 C ; 6 days at 2 - 8 C
o
Performance Characteristics Please consult local regulations for a correct waste disposal.
measured at a wavelength 492 nm, is directly proportional to S56: dispose of this material and its container at hazardous or
creatinine concentration in the sample.
o
6 months at -20 C away from light Precision
Within run (Repeatiblity) special waste collection point.
Alkaline pH S57: use appropriate container to avoid environmental
Creatinine + picrate yellow-red complex System Parameters Level 1 Level 2 contamination.
n 20 20 S61: avoid release in environment. refer to special instructions/
Reagents Wavelength 492 nm
safety data sheets.
Optical path 1 cm Mean (mg/dL) 1.55 4.58
Assay type Fixed Rate
Standard (ST)
Direction increase SD 0.069 0.1 References
2 mg/dL 177 mmol/L 1. Bowers LD, Wong ET: kinetic serum creatinine assays. II. A
Sample : Reagent Ratio 1 : 10 CV% 4.45 2.2
e.g.: Reagent volume 1 ml critical evaluation and review. Clin Chem 26:555, 1980.
Reagent 1 (R1) 2. Doolan PD, Alpen EL, Theil GB: A clinical appraisal of the
Sample volume 100 ml Run to run (Reproducibility)
Picric acid 25 mmol/L plasma concentration and endogenous clearence of creatinine.
First read time 30 seconds
Surfactants Level 1 Level 2 AM J Med 32:65, 1962.
delay time 120 seconds
Creatinine Picric Acid Reagent contains a low concentration of picric 3. DI Giorgio J: Nonprotein nitrogenous constituents. In: clinical
last read time 150 seconds n 20 20
acid, a chemical which, in its dry form, is flammable and potentially o chemistry – principles and technics, 2 nd ed. RJ Henry, DC
Temperature 25 C Mean (mg/dL) 1.67 4.63
explosive. For this reason, it is recommended that drains be well Cannon, JW Winkelman, editors, Harper and Row, Hagerstown
Zero adjustment Against Air
flushed with water when disposing the reagent, spills be cleaned up SD 0.081 0.19 (MD), 1974, pp 541-553.
at once, and avoid dryness of the material around the reagent bottle 4. Spencer K, Price CP: A review of Non-enzyme mediated
High 0.8 AU CV% 4.58 2.7
opening. reaction and their application to centrifugal analyzers.
Sensitivity 0.31 mg/dL (0.027 mmol/L)
Linearity 20 mg/dL (1.77 mmol/L) IN centerfugal analyzers in clinical chemistry, CP Price, K
Reagent 2 (R2) Methods Comparison Spencer, editors, Praeger publishers, New York, 1980, p231.
Sodium hydroxide 0.4 mol/L A comparison between Spectrum Diagnostics Creatinine Jaffè 5. Tobias GJ, Mclaughlin RF, Hopper J: Endogenous creatine
Irritant (xi) R36/38: Irritating to eyes and skin. Procedure reagent and a commercial reagent of the same methodology was clearence. N Engl j Med 266:317, 1962.
S26: In case of contact with eyes, rinse immediately with plenty of Pipette into test tubes performed on 20 human sera. A correlation of 0.991 was obtained. 6. Tietz NW: Textbook of clinical chemistry. WB saunders,
water and seek medical advice. philadelphia, 1986, pp 1271- 1281.
Working solution 1.0 ml
S37/39: Wear suitable gloves and eye/face protection. Sensitivity
Standard or Specimen 100 ml
When run as recommended, the minimum detection of this assay is
For further information, refer to the Creatinine Jaffè reagent material 0.31 mg/dL creatinine (0.027 mmol/L).
Mix, and after 30 seconds. read the absorbance A1 of the standard
safety data sheet. CATALOG NO QUANTITY
or specimen. After exactly 2 minutes later, read absorbance A2 of
standard or specimen. Linearity 234 001 2 x 100 ml
Precautions and Warnings The reaction is linear up to serum creatinine concentration of 20mg/ 234 002 4 x 100 ml
Do not ingest or inhalate. In case of contact with eyes or skin; rinse dL (1.77 mmol/L). Specimens showing higher concentration should 234 003 8 x 100 ml
immediately with plenty of soap and water. In case of severe injuries; be diluted 1+4 using physiological saline and repeat the assay 234 004 2 x 500 ml
seek medical advice immediately. (result×5). 234 005 2 x 250 ml
234 006 4 x 250 ml
30 31
Creatinine – Jaffè SYMBOLS IN PRODUCT LABELLING
Correction for body surface area can be done using the following
formula for creatinine clearance:
Expected Values
(Single Reagent) EC REP Authorised Representative

IVD For in-vitro diagnostic use !
CAUTION. Consult instructions Serum creatinine / min. per standard surface area =
Serum, plasma
Females 0.7-1.3 mg/dL 62-115 mmol/L
LOT Batch Code/Lot number for use Males 0.9-1.5 mg/dL 80-133mmol/L
REF: 237 001 (2 x 100 ml) 200 test UCr x V 1.73
REF: 237 002 (4 x 100 ml) 400 test REF Catalogue Number Manufactured by x
PCr A Urine(24 hrs)
Females 0.9 – 1.6 g/24 hrs
Where: UCr = Concentration of creatinine in urine( mg/dl) Males 1.1 – 2.8 g/24 hrs
Intended Use PCr = Concentration of creatinine in plasma(mg/dl)
Spectrum Diagnostics creatinine reagent is intended for the in-vitro V = Volume of urine flow in mL/min. Creatinine clearance
quantitative diagnostic determination of creatinine in human serum A = Body surface area in square meter . Females 75 – 115 ml / min.
Specimen Collection and Preservation
or urine on both automated and manual systems. 1.73/A = Factor normalizes clearance for average body Males 85 – 125 ml / min.
Serum or plasma
Both are suitable for analysis. The only acceptable anticoagulants surface.
Background are heparin and EDTA. Specimen should be promptly separated
Creatine is synthesized in kidney, liver and pancreas. It is Note: Body surface area can be determined from height weight Spectrum Diagnostics does not interpret the results of a
from cells after blood collection. The biological half-life of creatinine clinical laboratory procedure; interpretation of the results is
transported in blood to other organs such as muscle and brain where in blood is few minutes. via normograms in Tietz(6).
it is phosphorylated to phosphocreatine. Some free creatine in o o considered the responsibility of qualified medical personnel.
Stability: 7 day 2 - 8 C ; > 1 year at -20 C. All indications of clinical significance are supported by
muscle is converted to creatinine daily and the amount of creatinine
Quality Control literature references.
produced is proportional to muscle mass. In the absence of renal Urine Normal & abnormal commercial control serum of known
disease, excretion rate of creatinine in an individual is relatively Thymol or toluene may be used for urine preservation. To determine concentrations should be analyzed with each run.
constant. Therefore, measurement of creatinine clearance is useful creatinine concentration in urine, dilute 1 part sample with 49 parts Analytical Range
in detecting renal disease and estimating the extent of impairment isotonic saline prior to assay. Multiply result by 50 to compensate 0.31 – 20 mg/dL (0.027-1.77 mmol/L).
of renal function.Both serum creatinine and urea levels are elevated Performance Characteristics
for dilution. Precision
in patients with renal malfunction, especially decreased glomerular o
Stability: 2 days at 15 - 25 C ; 6 days at 2 - 8 C
o
Waste Disposal
filtration. In the early stage of kidney damage, increase in serum urea Within run (Repeatiblity)
o
6 months at -20 C away from light This product is made to be used in professional laboratories.
level usually precedes the increase in serum creatinine. However Level 1 Level 2 Please consult local regulations for a correct waste disposal.
serum urea levels may be affected by dehydration, diet and protein S56: dispose of this material and its container at hazardous or
System Parameters n 20 20
metabolism.On the other hand serum creatinine levels tend to be special waste collection point.
constant and unaffected by such factors. Thus serum creatinine is a Mean (mg/dL) 1.55 4.58 S57: use appropriate container to avoid environmental
Wavelength 492 nm
significantly more reliable renal function screening test than serum contamination.
Optical path 1 cm SD 0.069 0.1
urea. S61: avoid release in environment. refer to special instructions/
Assay type Fixed Rate
CV% 4.45 2.2 safety data sheets.
Direction increase
Method Sample : Reagent Ratio 1 : 10
Buffered Kinetic jaffé reaction without deproteinization. Run to run (Reproducibility) References
e.g.: Reagent volume 1 ml
Sample volume 100 ml Level 1 Level 2 1. Bowers LD, Wong ET: kinetic serum creatinine assays. II.
Assay Principle First read time 30 seconds A critical evaluation and review. Clin Chem 26:555, 1980.
Creatinine reacts with picric acid under alkaline condition to form n 20 20
delay time 120 seconds 2. Doolan PD, Alpen EL, Theil GB: A clinical appraisal of the
a yellow-red complex. The absorbance of the color produced, last read time 150 seconds Mean (mg/dL) 1.67 4.63 plasma concentration and endogenous clearence of creatinine.
measured at a wavelength 492 nm, is directly proportional to Temperature 30 C
o
AM J Med 32:65, 1962.
SD 0.081 0.19
creatinine concentration in the sample. Zero adjustment Against Air 3. DI Giorgio J: Nonprotein nitrogenous constituents. In: clinical
Reagent Blank Limits Low 0.30 AU CV% 4.58 2.7 chemistry – principles and technics, 2 nd ed. RJ Henry, DC
Alkaline pH
Creatinine + picrate yellow-red complex High 0.8 AU Cannon, JW Winkelman, editors, Harper and Row, Hagerstown
Sensitivity 0.31 mg/dL (0.027 mmol/L) Methods Comparison (MD), 1974, pp 541-553.
Reagents Linearity 20 mg/dL (1.77 mmol/L) A comparison between Spectrum Diagnostics Creatinine Jaffè Single 4. Spencer K, Price CP: A review of Non-enzyme mediated
Standard (ST) reagent and a commercial reagent of the same methodology was reaction and their application to centrifugal analyzers.
2 mg/dL 177 mmol/L Procedure performed on 20 human sera. A correlation of 0.991 was obtained. IN centerfugal analyzers in clinical chemistry, CP Price, K
Spencer, editors, Praeger publishers, New York, 1980, p231.
Reagent (R) Pipette into test tubes Sensitivity 5. Tobias GJ, Mclaughlin RF, Hopper J: Endogenous creatine
Picric acid 25 mmol/L Reagent (R) 1.0 ml When run as recommended, the minimum detection of this assay is clearence. N Engl j Med 266:317, 1962.
Sodium hydroxide 0.4 mol/L Standard or Specimen 100 ml 0.31 mg/dL creatinine (0.027 mmol/L). 6. Tietz NW: Textbook of clinical chemistry. WB saunders,
philadelphia, 1986, pp 1271- 1281.
Irritant (xi) R36/38: Irritating to eyes and skin. Mix, and after 30 seconds. read the absorbance A1 of the standard Linearity
S26: In case of contact with eyes, rinse immediately with plenty of or specimen. After exactly 2 minutes later, read absorbance A2 of The reaction is linear up to serum creatinine concentration of 20mg/ ORDERING INFORMATION
water and seek medical advice. standard or specimen. dL (1.77 mmol/L). Specimens showing higher concentration should
S37/39: Wear suitable gloves and eye/face protection. be diluted 1+4 using physiological saline and repeat the assay CATALOG NO QUANTITY
Calculation (result×5). 237 001 2 x 100 ml
For further information, refer to the Creatinine Jaffè reagent material A2 – A1 = Aspecimen or Astandard. 237 002 4 x 100 ml
safety data sheet. Interfering Substances 237 003 2 x 500 ml
Concentration of creatinine in serum: Serum, plasma
Precautions and Warnings (Aspecimen)
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Creatinine (mg/dL) = x2 Haemolysis
immediately with plenty of soap and water. In case of severe injuries; (Astandard)
Erythrocyte contamination doesn’t elevate results.
Concentration of creatinine in urine: Icterus
Reagent Preparation, Storage and Stability (Aspecimen) Serum bilirubin levels higher than 5 mg/dL (85 mmol/L) decrease
Spectrum Creatinine reagent is supplied ready-to-use Creatinine (mg/dL) = x 2 x 50 serum creatinine.
and stable up to the expiry date labeled on the bottles when (Astandard)
o
properly stored at 2 - 8 C . Lipemia
Creatinine clearance (ml/minutes): Lipemic specimens may cause high absorbance flagging. Diluted
Deterioration mg creatinine / dl urine x ml urine / 24 hours
sample treatment may be recommended.
The creatinine reagent are not suitable for use if combined reagents
have an absorbance greater than 0.8 at 492 nm measured in a 1cm mg creatinine / dl serum x 1440
lightpath or if the reagents develop a hazy appearance.
32 33
Creatinine - Colorimetric SYMBOLS IN PRODUCT LABELLING
Correction for body surface area can be done using the following
formula for creatinine clearance:
clinical laboratory procedure;interpretation of the results is
EC REP Authorised Representative Use by/Expiration Date
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions Serum creatinine / min. per standard surface area =
REF: 235 001 2 x 100 ml All indications of clinical significance are supported by
LOT Batch Code/Lot number for use literature references.
REF: 235 002 4 x 100 ml UCr x V 1.73
REF: 235 003 2 x 800 ml REF Catalogue Number Manufactured by x
Consult instructions for use (Xi) - Irritant PCr A
REF: 235 004 2 x 500 ml
Temperature Limitation (C) - Corrosive
Where: UCr = Concentration of creatinine in urine (mg/dL) Dynamic Range
PCr = Concentration of creatinine in plasma (mg/dL) 0.4 - 15 mg/dL (0.035 - 1.32 mmol/L).
V = Volume of urine flow in mL/min.
Intended Use
Reagent Storage and Stability A = Body surface area in square meter .
Waste Disposal
Spectrum Diagnostics creatinine reagent is intended for the in-vitro
All reagents are stable until expiration date stated on label when 1.73/A = Factor normalizes clearance for average body
quantitative, diagnostic determination of creatinine in human serum This product is made to be used in professional laboratories.
stored at 15 - 25 oC. surface.
or urine on manual system. Please consult local regulations for a correct waste disposal.
Working solution is stable for 5 hours at 15 – 25 oC away from light. Note : Body surface area can be determined from height weight
via normograms in Tietz(6). special waste collection point.
Background
Deterioration S57: use appropriate container to avoid environmental
Creatine is synthesized in kidney, liver and pancreas. It is
The creatinine reagents are not suitable for use if combined reagents Quality Control contamination.
transported in blood to other organs such as muscle and brain where
have an absorbance greater than 0.6 at 492 nm measured in a Normal & abnormal commercial control serum of known S61: avoid release in environment. refer to special instructions/
it is phosphorylated to phosphocreatine. Some free creatine in
1-cm lightpath or if the reagents develop a hazy appearance. concentrations should be analyzed with each run. safety data sheets.
muscle is converted to creatinine daily, and the amount of creatinine
produced is proportional to muscle mass. In the absence of renal
disease, excretion rate of creatinine in an individual is relatively Methods Comparison
A comparison between Spectrum Diagnostics Creatinine colorimetric References
constant. Therefore, measurement of creatinine clearance is useful
Specimen Collection and Preservation reagent and acommercial reagent of the same methodology was 1. Bowers LD, Wong ET: kinetic serum creatinine assays. II. A
in detecting renal disease and estimating the extent of impairment of
performed on 40 human sera. A correlation (r) of 0.996 was obtained. critical evaluation and review. Clin Chem 26:555, 1980.
renal function. Both serum creatinine and urea levels are elevated
Serum or plasma 2. DI Giorgio J: Nonprotein nitrogenous constituents. In :clinical
in patients with renal malfunction, especially decreased glomerular
chemistry – principles and technics, 2 nd ed.RJ Henry, DC
filtration. In the erarly stage of kidney damage, increase in serum Both are suitable for analysis. The only acceptable anticoagulants Sensitivity
are heparin and EDTA. Specimen should be promptly separated Cannon, JW Winkelman, editors, Harperand Row, Hagerstown
urea level usually precedes the increase in serum creatinine When run as recommended, the minimum detection limit of the
from cells after blood collection. The biological half-life of creatinine (MD), 1974, pp 541-553.
.However serum urea levels may be affected by dehydration, diet assay is 0.4 mg/dL (0.035 mmol/L).
in blood is few minutes. 3. Doolan PD, Alpen EL, Theil GB: A clinical appraisal of the
and protein metabolism. On the other hand serum creatinine levels
Stability: 7 days 2 - 8 oC ; > 1 year at – 20 oC plasma concentration and endogenous clearence of creatinine.
tend to be constant and unaffected by such factors. Thus serum Linearity AM J Med 32:65, 1962.
creatinine is a significantly more reliable renal function screening The reaction is linear up to a creatinine concentration of 15 mg/dL;
4. Spencer K, Price CP: A review of Non-enzyme mediated
test than serum urea. Urine specimens showing higher concentration should be diluted 1+4
reaction and their application to centrifugal analyzers. In:
Thymol or toluene may be used for urine preservation. To determine using physiological saline and repeat the assay (result × 5).
Centerfugal analyzers in clinical chemistry , CP Price, K
Method creatinine concentration in urine, dilute 1 part sample with 49 parts
Spencer, editors, Praeger publishers, New York, 1980, p 231.
Colorimetric method with deproteinization. isotonic saline prior to assay. Multiply result by 50 to compensate Interfering Substances 5. Tobias GJ, Mclaughlin RF, Hopper J: Endogenous creatine
for dilution. Serum, plasma
o o clearence. N Engl J Med 266:317, 1962.
Assay Principle Stability: 2 days at 15 - 25 C ; 6 days at 2 - 8 C
o 6. Tietz NW: Textbook of clinical chemistry. WB saunders,
6 months at -20 C away from light Haemolysis philadelphia, 1986, pp 1271- 1281.
Reagents Erythrocyte contamination doesn’t elevate results.
Standard creatinine (ST) Procedure
2 mg/dL 177 mmol/L Pipette into test tubes Icterus ORDERING INFORMATION
Blank Standard Sample Urine Serum bilirubin levels in the pathological range may interfere with CATALOG NO QUANTITY
Reagent 1 (R1) the results.
Distilled Water 0.5 ml ------- ------- ------- 235 001 2 x 100 ml
Picric acid 38 mmol/L
Lipemia 235 002 4 x 100 ml
Reagent 1 contains a low concentration of picric acid, a chemical Standard ------- 0.5 ml ------- -------
Lipemic specimens may cause high absorbance flagging. Diluted 235 003 8 x 100 ml
which, in its dry form, is flammable and potentially explosive. For
TCA 0.5 ml 0.5 ml ------- 0.5 ml sample treatment may be recommended. 235 004 2 x 500 ml
this reason, it is recommended that drains be well flushed with water
when discarding the reagent, spills be cleaned up at once, and dried Supernatant ------- ------- 1.0 ml -------
material not be allowed to build up around the reagent bottle opening Urine (1+ 49) ------- ------- ------- 0.5 ml
Expected Values
Serum, plasma
Reagent mixture 1.0 ml 1.0 ml 1.0 ml 1.0 ml Females 0.7-1.3 mg/dL 62-115 mmol/L
Reagent 2 (R2)
Sodium hydroxide 1.6 mol/L Males 0.9-1.5 mg/dL 80-133 mmol/L
o
Mix and let stand for 20 minutes. at 20–25 C. Measure the
Reagent 2 contains caustic material.
absorbance of specimen and standard against reagent blank at Urine(24 hrs)
Corrosive (C) 546 nm. Females 0.9 – 1.6 g/24 hrs
R35 cause severe burns.
Males 1.1 – 2.8 g/24 hrs
R41 Risk of serious damage to eyes.
Calculation
S26 In case of contact with eyes, rinse immediately with
Concentration of creatinine in serum: Creatinine clearance
(Aspecimen) Females 75 – 115 mL / min
S28 After contact with skin, wash immediately with plenty of
Creatinine (mg/dL) = x 2 Males 85 – 125 mL / min
soap and water. (Astandard)
Reagent Preparation
Prepare working solution as following: Concentration of creatinine in urine:
Combine one volume of R1 with one volume of
R2, e.g. 1.0 ml R1 + 1.0 ml R2 (Aspecimen)
Creatinine (mg/dL) = x 2 x 50
(Astandard)
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Creatinine clearance:
mg creatinine / dL urine x mL urine / 24 hours
mg creatinine / dL serum x 1440
34 35
GLUCOSE - Liquizyme SYMBOLS IN PRODUCT LABELLING
Calculation
Glucose concentration (mg/dl) =
(Aspecimen)
× 100
Urine
Random 5.0 - 15 mg/dL (0.28 - 0.83 mmol/L)
GOD - PAP (Single Reagent) (Astandard) 24 hours < 0.5 g/24 hrs (<2.8 mmol/24 hrs)
EC REP Authorised Representative Temperature Limitation
IVD For in-vitro diagnostic use Use by/Expiration Date Quality Control
REF: 250 001 (2 x 100 ml) 200 test CSF
LOT Batch Code/Lot number CAUTION. Consult instructions Normal & abnormal commercial control serum of known
REF: 250 002 (4 x 100 ml) 400 test Adults 40 - 75 mg/dL (2.2-4.2 mmol/L)
REF Catalogue Number for use concentrations should be analyzed with each run.
REF: 250 003 (8 x 100 ml) 800 test
REF: 250 004 (2 x 500 ml) 1000 test Consult instructions for use Manufactured by
Performance Characteristics CSF glucose values should be approximately 60% of the plasma
REF: 250 005 (2 x 250 ml) 500 test values and must always be compared with concurrently measured
REF: 250 006 (4 x 250 ml) 1000 test Precision
Reagent Preparation, Storage and Stability plasma values for adequate clinical interpretation.
REF: 250 007 (1 x 250 ml) 250 test Within run (Repeatiblity)
Spectrum Glucose liquizyme reagents are supplied ready-to-use
and stable up to the expiry date labeled on the bottles when properly Level 1 Level 2
Intended Use o
stored refrigerated at 2 – 8 C. Once opened, the opened vial is n 20 20 clinical laboratory procedure;interpretation of the results is
Spectrum Diagnostics liquizyme glucose reagent is intended for the stable for 3 months at the specified temperature.
in-vitro quantitative, diagnostic determination of glucose in human Mean (mg/dL) 103 228
serum, plasma, urine and CSF on both manual and automated Deterioration SD 1.12 1.19 literature references.
systems. The Spectrum glucose reagent is normally clear or pale pink. Do not
use liquizyme Glucose reagent if it is turbid or if the absorbance is CV% 1.09 0.83
Background greater than 0.2 at 546 nm. Dynamic Range
Oxidation of glucose present in the peripheral blood represents Run to run (Reproducibility) 5 - 500 mg/dL (0.27 - 27.7 mmol/L).
the major source of cellular energy in the body. Dietary glucose is Specimen Collection and Preservation Level 1 Level 2
stored in the liver in the form of glycogen or converted to fatty acids Waste Disposal
and stored in the adipose tissues. The accurate estimation of glucose n 20 20
Serum or plasma This product is made to be used in professional laboratories.
is important in the diagnosis and management of hyperglycemia Individuals should be fasting before sample collection.Heparin, Mean (mg/dL) 109 235 Please consult local regulations for a correct waste disposal.
& hypoglycemia. the most frequent cause of hyperglycemia is EDTA, and flouride are the only accepted anticoagulants The stability S56: dispose of this material and its container at hazardous or
SD 1.23 1.27
diabetes mellitus resulting from a deficiency in insulin secretion or of glucose in specimen is affected by storage temperature, bacterial special waste collection point.
action. Hypoglycemia may be the result of an insulinoma, insuline contamination and glycolysis. Serum or plasma should be separated CV% 1.17 0.98 S57: use appropriate container to avoid environmental
administration, inborn error of carbohydrate metabolism or fasting. within 30 minutes.when blood is drawn and permitted to clot and to contamination.
The concentration of glucose in the blood is controlled within narrow stand uncentrifuged at room temperature. The average decrease in Methods Comparison S61: avoid release in environment. refer to special instructions/
limits by many hormones, the most important of which are produced serum glucose is 7% in 1 hour (0.28 to 0.56 mmol/l or 5 to 10 mg/ A comparison between Spectrum Diagnostics Glucose reagent and safety data sheets.
by the pancreas. dl). This decrease is the result of glycolysis. Unhemolyzed serum a commercial reagent of the same methodology was performed on
GLUCOSE measurement in urine is used as diabetes screening o o
glucose is stable up to 8 hours at 25 C or up to 72 hours at 4 C. 20 human sera. A correlation of 0.991 was obtained. References
procedure and to aid in the evaluation of glucosuria to detect renal In order to inhibit glycolysis, samples should be collected into tubes 1. Caraway WT, Watts NB. Carbohydrates In : Tietz NW,
tubular defect and in the management of diabetes mellitus. containing sodium fluoride. Sensitivity ed.Fundamentals of Clinical Chemistry. 3ry ed. Philadephia
GLUCOSE measurement in cerebrospinal fluid (CSF) is used for Urine When run as recommended, the minimum detection limit of the WB saunde-rs 1987:422-447.
evaluation of meningitis, neoplastic involvement of meninges and o
Urine samples are stable 1 day at 4 C , in case of delay due to assay is 5 mg/dL (0.27 mmol/L). 2. Howanitz PJ, Howanitz JH. Carbohydrates. In: Henry JB,ed.
other neurological disorders. transportation or for 24 hour urine collection, it is recommended to Clinical diagnosis and mana-Gement by laboratory methods.
add either merthiolate (0.23 mmol/L) or 5 ml glacial acetic acid to Linearity 17th ed Philadelphia: WB saunders 1984:165-179
Method the container before collection. Unpreserved urine samples may The reaction is linear up to glucose concentration of 500 mg/dl; 3. Trinder, P., Ann. Clin. Biochem. (1969), 6:24.
GOD-PAP enzymtic colorimetric method. lose up to 40% of their glucose after 24 hour storage at room specimens showing higher concentration should be diluted 1+2 4. Tietz NW, ed. Clinical guide to laboratory tests. 3rd
temperature; therefore, keep samples on ice during collection. using physiological saline and repeat the assay (result×3). ed.Philadelphia: WB saunders; 1995:268-273.
Assay Principle CSF 5. Weissman M, klien B. Evaluation, of glucose determination In
Glucose is determined after enzymatic oxidation in the presence Sample should be analyzed for glucose immediately to avoid Interfering Substances untreated serum samples. Clin Chem.1958;4:420-422.
of glucose oxidase. The formed hydrogen peroxide reacts under contamination with bacteria. If a delay in measurement is Serum, plasma
o
catalysis of peroxidase (PAP) with phenol and 4-aminoantipyrine to unavoidable, the sample should be centrifuged and stored at 4 C. ORDERING INFORMATION
form a red violet quinoneimine dye as indicator. Haemolysis
System Parameters No significant interference from haemoglobin up to 500 mg/dL. CATALOG NO QUANTITY
Glucose Gluconic acid
GOD + Wavelength 546 nm (492 – 550 nm) 250 001 2 x 100 ml
+ Optical path 1 cm
2H2O + O2 H2O2 lcterus 250 002 4 x 100 ml
Assay type End-point No significant interference from free and conjugated bilirubin up 250 003 8 x 100 ml
Direction Increase to levels of 15 mg/dL (257 mmol/L). 250 004 2 x 500 ml
2H2O2 +Phenol 4H2O
PAP + Sample : Reagent Ratio 1 : 100 250 005 2 x 250 ml
+
4-amino-antipyrine Quinoneimine e.g.: Reagent volume 1 ml lipemias 250 006 4 x 250 ml
Sample volume 10 ml Lipid disturb measurements if present in high concentration 250 007 1 x 250 ml
o o
Reagents Temperature 37 C or 20 – 25 C (More than 500 mg/dL).
o
Glucose standard (St) 100 mg/dL Incubation time 20 minutes at 20 – 25 C or
o
5.55 mmol/L 10 minutes at 37 C Others
Reagent (R) Zero adjustment Reagent Blank Turbidity caused by insoluble uranyl phosphate may result in false
Phosphate Buffer 100 mmol/L Reagent Blank Limits Low 0.00 AU high levels.
Phenol 4.0 mmol/L High 0.15 AU
4-amino-antipyrine 1.0 mmol/L Sensitivity 5 mg/dL (0.27 mmol/L) Reducing Substances
Glucose oxidase > 20 KU /L Linearity 500 mg/dL ( 27.7 mmol/L) Large amounts of reducing substances as ascorbic acid,
Peroxidase > 2.0 KU/L creatinine,glutathione and uric acid react with hydrogen peroxide
Sodium Azide 8 mmol/L Procedure and stimulate low glucose concentration.
Blank Standard specimen
For further information, refer to the Glucose reagent material safety Expected Values
data sheet. Reagent (R) 1.0 ml 1.0 ml 1.0 ml Serum, plasma
Standard ------- 10 ml ------- Adults (fasting) 70 - 105 mg/dL (3.9 -5.8 mmol/L)
Precautions and Warnings Children 60 - 110 mg/dL (3.33-6.11 mmol/L)
Specimen ------- ------- 10 ml Newborns 40 - 60 mg/dL (2.22 – 3.33 mmol/L)
immediately with plenty of soap and water. In case of severe injuries; o
Mix and incubate for 10 minutes at 37 C or 20 minutes at
15 -25 C. Measure absorbance of specimen (Aspecimen) and
o
Reagent (R) contains sodium azide which may react with copper or standard (Astandard) against reagent blank within 30 minutes.
lead plumbing.
36 37
GLYCOSYLATED HEMOGLOBIN (GHb) Calculations
(Ion Exchange Resin method)
For the quantitative determination of Glycohemoglobin in Blood Abs. Control GHb
Ratio of Control (RC) = ——————————
REF: 254 001 25 Tests Abs. Control THb
REF: 254 002 50 Tests
REF: 254 003 100 Tests Abs. Test GHb
Ratio of Test (RT) = ——————————
Abs. Test THb
Background

Glycosylated hemoglobin (GHb) is formed continuously by the adduction of glucose by co-valent bonding to the amino-terminal valine of the
Ratio of Test (RT)
hemoglobin beta chain progressively & irreversibly over a period of time & is stable till the life of the RBC. This process is slow, non enzymatic
GHb in % = ————————— x 10 (Value of Control)
and is dependent on the average blood Glucose concentration over a period of time.
Ratio of Control (RC)
A single glucose determination reflects the glucose level at that time. GHb on the other hand reflects the mean glucose level over an
extended period of time. Thus GHb reflects the metabolic control of glucose level over a period of time unaffected by diet, insulin, other
drugs, or exercise on the day of testing. GHb is now widely recognized as an important test for the diagnosis of Diabetes mellitus and is a Linearity
reliable indicator of the efficacy of therapy. The Glycosylated hemoglobin procedure shows linearity for GHb levels in the range of 4.0% - 20.0%.
Assay Principle Notes

Glycosylated hemoglobin (GHb) has been defined operationally as the fast fraction hemoglobins HbA1 (Hb A1a, A1b, A1c) which elute Blood samples with Hemoglobin greater than 18 g/dl should be diluted 1 + 1 with Normal saline before the assay.
first during column chromatography. The non - glycosylated hemoglobin, which consists of the bulk of hemoglobin, has been designated Samples from patients with Hemoglobinopathies, decreased red cell survival times, gross lipemia may show incorrect results.
HbAo. A hemolysed preparation of whole blood is mixed continuously for 5 minutes with a weakly binding cation-exchange resin. The labile Do not use Ion Exchange Resin tubes in case of turbidity or visible discolouration. Diabetics with metabolic imbalance may have extremely
fraction is eliminated during the hemolysate preparation and during the binding. During this mixing, HbAo binds to the ion exchange resin high levels of the labile aldimine form. In such cases the incubation time during hemolysate preparation may be increased to 15 minutes to
leaving GHb free in the supernatant. After the mixing period, a filter separator is used to remove the resin from the supernatant. The percent ensure elimination of this in stable fraction.
glycosylated hemoglobin is determined by measuring absorbances of the ratio of the absorbances of the Glycosylated hemoglobin (Ghb) &
the Total hemoglobin fraction (THb) . The ratio of the absorbances of GHb and THb of the control and test is used to calculate the percent References
GHb of the sample.
1. Trivelli,L.A.,Ranney,H.M. and Lai,H.T., New Eng.J.Med. 284, 353 (1971).
Expected values 2. Nathan,D.M.,et al., New Eng.J.Med. 310, 341 - 346 (1984).
Normal < 8.0 %
Good control 8.0 – 9.0 % 3. Bunn,H.F., Diabetes 130, 613 (1981).
Fair control 9.0 – 10.0 %
4. Bates,H.M.,Lab Manag.,Vol 16 ( Jan.1978)
Poor control > 10.0 %
It is recommended that each laboratory establish its own normal range representing its patient population.
Reagents 25 Tests 50 Tests 100 Tests

R1 Ion Exchange Resin (Predispensed Tubes) 25 x 3 ml 50 x 3 ml 100 x 3 ml
R2 Lysing Reagent 1 x 12.5 ml 2 x 12.5 ml 4 X 12.5 ml
C Control (10% GHb) 1 x 1 ml 2 x 1 ml 4 x 1 ml
A Resin Separators 25 Nos 50 Nos 100 Nos

Reagents Preparation Storage and Stability
Contents are stable at 2-8°C till the expiry mentioned on the label. Do not freeze. The Resin separators can be removed on opening the kit
and stored at R.T.
The Ion Exchange Resin tubes & the Lysing Reagent are ready to use. Reconstitute the Control with 1 ml of distilled water. Allow to stand
for 10 mins with occasional mixing. The reconstituted control is stable for at least 7 days when stored at 2-8°C tightly sealed, and at least 4
weeks when stored at -20°C. Do not thaw and refreeze.
Specimens: Whole blood, preferably fresh collected in EDTA. GHb in whole blood is stable for week at 2-8°C
Procedure
Wavelength : 415 nm (Hg 405 nm)
Temperature : R.T.
Light path : 1 cm
A. Hemolysate Preparation
1. Dispense 0.5 ml Lysing Reagent into tubes labeled as Control (C) & Test (T).
2. Add 0.1ml of the reconstituted control & well-mixed blood sample into the appropriately labeled tubes. Mix until complete lysis is evident.
3. Allow to stand for 5 minutes.
B. Glycosylated hemoglobin (GHb) Separation

Remove cap from the Ion-Exchange Resin tubes and label as Control & Test.
Add 0.1 ml of the hemolysate from Step A into the appropriately labeled Ion Exchange Resin tubes.
Insert a resin Separator into each tube so that the rubber sleeve is approximately 1 cm above the liquid level of the resin suspension.
Mix the tubes on a rocker, rotator or a vortex mixer continuously for 5 minutes.
Allow the resin to settle, then push the resin separator into the tubes until the resin is firmly packed.
Pour or aspirate each supernatant directly into a cuvette and measure each absorbance against distilled water.
C. Total Hemoglobin (THb) fraction

Dispense 5.0 ml of distilled water into tubes labeled as Control & Test.
Add to it 0.02 ml of hemolysate from Step A into the appropriately labeled tube.
Mix well.
Read each absorbance against distilled water
38 39
Direct Enzymatic HbA1c SYMBOLS IN PRODUCT LABELLING
EXPECTED VALUES
Recommended Values are
REFERENCES
1. Trivelli, L.A., Ranney, H.M., and Lai, H.T., New Eng. J. Med.
EC REP Authorised Representative Use by/Expiration Date < 6% for non-diabetics (42 mmol/mol) 284,353 (1971).
REF: 255 001 60 Tests IVD For in-vitro diagnostic use ! CAUTION. Consult instructions 6 - 9% (42 – 75 mmol/mol) for diabetics under 2. Gonen, B., and Rubenstein, A.H., Diabetologia 15, 1 (1978).
LOT Batch Code/Lot number for use perfect glycemic control
R1a: 1 x 7 ml Up to 20% for diabetics out of glycemic control 3. Gabbay, K.H., Hasty, K., Breslow, J.L., Ellison, R.C., Bunn, H.F.,
R1b: 1 x 3 ml and Gallop, P.M., J. Clin. Endocrinol. Metab. 44, 859 (1977).
Consult instructions for use (Xn) - Harmful
R2: 1 x 4.4 ml Temperature Limitation Conversion from HbA1C % to mmol/mol 4. Bates, H.M., Lab. Mang., Vol 16 (Jan. 1978).
LR: 1 x 13 ml 5. Tietz, N.W., Textbook of Clinical Chemistry, Philadelphia, W.B.
HbA1c % HbA1C mmol/mol Saunders Company, p.794-795 (1999).
6.0 42 6. Corielo, A., et al, Diabetologia 22, p. 379 (1962).
7. Goldstein, D.E., et al, Clin. Chem. 32, pp. 364-370 (1986).
6.5 48
8. Fluckiger, R., et al, Med Intelligence 304 pp. 823-827 (1981).
7.0 53
9. Nathan, D.M., et al, Clin. Chem. 29, pp. 466-469 (1983).
GENERAL 7.5 59 10. American Diabetes Association: Clinical Practice
SAMPLES
The glycemic control in diabetes mellitus is performed mainly by the Collect venous blood with EDTA. 8.0 65 Recommendations, Diabetes Care 24 (Suppl. 1):
determination of Glucose, but also through quantitative determination Storage and Stability
of Hemoglobin A1c (HbA1c) in human blood : HbA1c is an indication 9.0 75
Hemoglobin A1c in whole blood with EDTA
for the actual glucose levels over the preceding 3 months. It was is stable for one week at 2-8°C.5 10 86
shown that HbA1c in diabetic subjects can be elevated 2-3 fold over ORDERING INFORMATION
normal and on the other hand approaches normal values when they
are under metabolic control. CATALOG NO QUANTITY
Hemolysate Procedure: Note: 255 000 30 Test
1. Dispense 250µl of Lysing Reagent into cups or tubes and label
PRINCIPLE Each laboratory should establish its own expected values. The given 255 001 60 Test
as Controls, Patients, etc.
Hemolyzed blood is used as sample material. Through protease values can only be an average indication.
2. Add 20ul of well mixed (!) whole blood samples respectively of
attack glycated valines are released and are subject to further Calibrators and Controls . (Note: Calibrators and Controls have
enymatic reaction with fructosyl valine oxidase (FVO). The result to be treated exactly like the patient samples!)
LIMITATIONS
is a measureable quantitative amount of hydrogen peroxide, which 1. Results may be inconsistent in patients e.g.with opiate
3. Let incubate at room temperature for minimum 10 min.
is determined through colorimetric reaction with a chromogen addiction, lead-poisoning, alcoholism, ingestion of large doses
Stability:
compound. of aspirin.
Hemolysates may be stored up to 1 day at 2-8°C
The reaction product is proportional to the amount of HbA1c and 2. Elevated levels( > 10%) of HbF may lead to underestimation
is measured as absorbance A. The HbA1c value is derived from a of HA1c.
calibration curve. PRECAUTIONS 3. Hemoglobin variants HbS, HbC and HbE do not interfere in this
1. 1.The reagent is for in vitro diagnostic use only. assay. There is also no interference by labile intermediates ,
2. 2.All human specimens should be regarded as potentially and uremia does not interfere, too.
REAGENTS biohazardous. Therefore, universal precautions should be used
R1a: MES-Buffer (pH 7.0) 5.0 mmol/l in specimen handling (gloves, lab garments, avoid aerosol PERFORMANCE CHARACTERISTICS
Protease production, etc.) 1. Linearity: The Hemoglobin A1c assay range is 2.0% to
Triton-X 100 0.5 %l 16.0%. Results in this range can be reported and used directly.
ANALYTICAL PROCEDURE
R1b: MES-Buffer (pH 6.3) 1.0 mmol/l Wavelength 700 nm 2. Correlation: A study using 40 human specimens between
Cuvette 1 cm light path this HA1c procedure and the reference method yielded
R2: TRIS-Buffer (pH 8.0) 5.0 mmol/l Temperature 37 °C a correlation coefficient of 0.9874 and a linear regression
Fructosyl Valine Oxidase ≥ 9,5 kU/l Specimen Blood equation of y = 1.021 x + 0.014
Peroxidase ≥ 8,5 kU/l
Chromogen ≥ 0.7mmol/l
Hemosylate Hemosylate 3. Precision:
LR: Lysing Reagent Calibrator Sample Within Run: The intra assay precision was established by
assaying blood with two Hemoglobin A1c levels twenty times
R1 (μL) 200 200 each.
Storage and Stability
Calibrator(μL) 30 -
Store all reagents refrigerated at 2-8°C. Unopened reagents are Level Mean % C.V.
stable up to the expiration date printed on the labels. Sample (μL) - 30 Medium 5.7 1.0
Mix and incubate for 5 minutes exactly, then Read the High 10.3 0.7
Preparation of Reagents absorbance (A1) of the samples and calibrator,
3-reagent-procedure: and immediately add R2
R1a, R1b and R2 are ready for use when the 3-reagent- INTERFERENCES
R2(μL) 85 85
procedure is applied. 1. Bilirubin to 15mg/dL, ascorbic acid to 10mg/dL, triglycerides
Stability after opening : Read the absorbance (A2) of the Samples and calibrator after
2 mins. to 3000mg/dL, Glucose to 4000mg/dL, carbamylated Hb to
At least 3 months when contamination is avoided 5mmol/L and acetylated Hb to 5.0mmol/L do not interfere in
CALCULATION the assay.
2-reagent-procedure: 2. It has been reported that results may be inconsistent in patients
R1: Mix 7 parts R1a with 3 parts R1b (D A) Sample
Stability: who have the conditions like opiate addiction, lead-poisoning,
────────── X Calibrator conc. = % HbA1c
At least 2 weeks when contamination is avoided (D A) Calibrator alcoholism, ingestion of large doses of aspirin.
Important Note: QUALITY CONTROL

R1b and R2 are light sensitive ! Use Spectrum HbA1C Control Set with assayed values.
Additional Reagents
Control Set.
Calibrator.
40 41
Haemoglobin(Single Reagent) SYMBOLS IN PRODUCT LABELLING Within run (Repeatiblity)
CATALOG NO QUANTITY
Drabkin’s Solution EC REP Authorised Representative Use by/Expiration Date Level 1 Level 2
610 001 2 x 20 ml
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions 610 002 5 x 20 ml
Batch Code/Lot number for use
n 20 20
LOT
REF:610 001 (2 x 20 ml) 800 test
REF Catalogue Number Manufactured by Mean (mg/dL) 10 14
REF:610 002 (5 x 20 ml) 2000 test
Consult instructions for use (Xn) - Harmful CV% 2.3 1.3
Run to run (Reproducibility)

Intended Use Reagent Preparation Storage and Stability Level 1 Level 2
Spectrum Diagnostics haemoglobin reagent is intended for the in- The reagent is stable until expiration date stated on label when stored n 20 20
o
vitro quantitative, diagnostic determination of haemoglobin in human at 15 - 25 C. Prepare the working solution by diluting the contents of
Mean (mg/dL) 11.1 14.1
blood. the contcentrated reagent (R) to 1000ml with distilled water (add one
CV% 2.9 2.1
bottle of reagent (R) to 980 ml of dist.water), mix thoroughly. working
Background reagent can be prepared according to the volume needed by mixing
Haemoglobin (Hb) is the red pigmented protein located in the 1 ml of the concentrated reagent (R) plus 49 ml distilled water. Spectrum Diagnostics does not interpret the results of a
er-ythrocytes and consists of four subunits. Its main function is clinical laboratory procedure; interpretation of the results is
the transport of oxygen and carbon dioxide in blood. In normal Stability: 6 months in a brown glass bottle at 15-30 C
o considered the responsibility of qualified medical personnel.
humanadults, at least 96 % of the haemoglobin is HbA. HbA2 is
usually about 2.5 – 3 % of total haemoglobin. Fetal hemoglobin (HbF) Specimen Collection and Preservation
predominates during fetal life and diminishes rapidly during the first Anticoagulated venous or capillary blood . Blood may be anticoa-
year of postnatal life. In normal adults less than 1 % is HbF.Blood gulated with EDTA, or fluoride. Blood can be taken directly from a Methods Comparison
haemoglobin concentration may be diminished as a consequence of finger or heel puncture without use of anticoagulant. A comparison between Spectrum Diagnostics Haemoglobin reagent
hemorrhage or hemolysis or as a result of impaired blood formation and a commercial reagent of the same methodology was performed
o
in the bone marrow. Stability : 7 days at 2 – 8 C on 40 human blood samples. A correlation (r) of 0.983 was obtained.
o
4 days at 20 - 25 C
Method
colorimetric method using Drabkin’s solution. System Parameters Expected values
Wavelength 540 nm (Hg 546 nm) 1- 6 days 15.2 – 23.5 g/dL ( 9.4 – 14.6 mmol/L)
Assay Principle Optical path 1 cm 14 – 50 days 10.3 – 16.6 g/dL ( 6.4 – 10.3 mmol/L)
Haemoglobin is oxidized by potassium ferricyanide which is Assay type End-point
2 - 10 months 10.0 – 12.9 g/dL ( 6.1 – 8.0 mmol/L)
converted into stable cyanomethaemoglobin by potassium cyanide. Direction Increase
1 – 15 years 11.0 – 14.3 g/dL ( 6.8 – 8.8 mmol/L)
The absorbance of the cyanomethaemoglobin is monitored at 540 Sample : Reagent Ratio 1 : 250
nm. e.g .: Reagent volume 2.5 ml Adults Women 12.0 – 16.0 g/dL (7.5 – 9.9 mmol/L)
Sample volume 10 ml Men 14.0 – 18.0 g/dL (8.7 – 11.2 mmol/L)
Reagents Temperature
o
20 – 25 C
Incubation time 5 minutes Waste Disposal
Reagent Zero adjustment Against reagent blank This product is made to be used in professional laboratories.
Potassium ferricyanide 31 mmol/l Please consult local regulations for a correct waste disposal.
Potassium phosphate 52 mmol/l Procedure S56: dispose of this material and its container at hazardous or
Potassium cyanide 77 mmol/l Pipette into test tubes special waste collection point.
Surfactant 2% working solution 2.5 ml S57: use appropriate container to avoid environmental
Harmful (Xn): R20/21/22: Harmful by inhalation, in contact with skin Blood sample 10 ml contamination.
and if swallowed. S7: Keep container tightly closed. S28.1: After S61: avoid release in environment. refer to special instructions/
contact with skin, wash immediately with plenty of water. Mix well and rinse the blood pipette several times with the reagents, safety data sheets.
o
S45: In case of accident or if you feel unwell, seek medical advice and incubate for 5 minuts at 20-25 C. Measure absorbance of
immediately. The amount of cyanide present in one bottle of reagent specimen (Aspecimen) against reagent blank.
References
is apreciably less than the minimum lethal dose for an adult.
1. International committee for standardization in haematology.
However, hydrogen cyanide is liberated by acidification. Never allow Calculation
Brit.J. Haemat., 1967:13 (Suppl.) 71.
reagent to come in contact with acid. Haemoglobin concentration (g/dL) = Aspecimen x 36.77
2. Van Kampen, E. J. and Zijlstra, W.G., Clin. Chem. Acta.,
1961:6:538 – 544.
For further information, refer to the Haemoglobin reagent material Haemoglobin concentration (mmol/L)= Aspecimen x 22.83
safety data sheet. 3. Tietz NW, Ed. Clinical guide to laboratory tests. 2ND ED.
Philadelphia: WB Saunders; 1990:566.
Quality Control
Precautions and Warnings Normal & abnormal commerical control blood of known ORDERING INFORMATION
Do not ingest or inhalate. In case of contact with eyes or skin; rinse concentrations should be analyzed with each run.
Precision
42 43
Haemoglobin SYMBOLS IN PRODUCT LABELLING
Drabkin’s Solution EC REP Authorised Representative

CAUTION. Consult instructions
Expected values
1- 6 days 15.2 – 23.5 g/dL ( 9.4 – 14.6 mmol/L)
LOT Batch Code/Lot number for use
REF Catalogue Number Manufactured by 14 – 50 days 10.3 – 16.6 g/dL ( 6.4 – 10.3 mmol/L)
REF: 610 003 (2 x 50 ml) 1000 test
Consult instructions for use (Xn) - Harmful 2 - 10 months 10.0 – 12.9 g/dL ( 6.1 – 8.0 mmol/L)
1 – 15 years 11.0 – 14.3 g/dL ( 6.8 – 8.8 mmol/L)
Adults Women 12.0 – 16.0 g/dL (7.5 – 9.9 mmol/L)
Men 14.0 – 18.0 g/dL (8.7 – 11.2 mmol/L)
Intended Use Precautions and Warnings Spectrum Diagnostics does not interpret the results of a
Spectrum Diagnostics haemoglobin reagent is intended for the Pay attention to all precautions and warnings listed in Spectrum clinical laboratory procedure ; interpretation of the results is
in-vitro quantitative, diagnostic determination of haemoglobin in Diagnostics catalogue available upon request .Haemoglobin reagent considered the responsibility of qualified medical personnel
human blood . contains cyanide which is poisonous. Avoid contact with skin and
literature references .
never pipette by mouth .
Background
Haemoglobin (Hb) is the red pigmented protein located in the Reagent Preparation Storage and Stability References
erythrocytes and consists of four subunits . Its main function is the All reagents are stable until expiration date stated on label when
o 1. International committee for standardization in haematology.
transport of oxygen and carbon dioxide in blood . In normal human stored at 15 - 25 C . Prepare the working solution by
Brit. J. Haemat., 1967:13 (Suppl.) 71 .
adults , at least 96 % of the haemoglobin is HbA . HbA2 is usually diluting the reagents with bidistilled water as following :
about 2.5 – 3 % of total haemoglobin . Fetal haemoglobin (HbF) 1 ml(R1) + 1 ml (R2) + 48 ml H2O 2. Van Kampen, E. J. and Zijlstra, W.G., Clin. Chem
Acta.,1961:6:538 – 544 .
predominates during fetal life and diminishes rapidly during the first working solution is stable for 3 months and should be light
year of postnatal life . In normal adults less than 1 % is HbF.Blood protected . 3. Tietz NW, Ed. Clinical guide to laboratory tests. 2ND ED.
Philadelphia: WB Saunders; 1990:566 .
haemoglobin concentration may be diminished as a consequence
of haemorrhage or haemolysis or as a result of impaired blood Specimen Collection and Preservation
formation in the bone marrow . Anticoagulated venous or capillary blood . Blood may be
anticoagulated with EDTA , or fluoride . Blood can be taken
Method directly from a finger or heel puncture without use of ORDERING INFORMATION
colorimetric method using Drabkin’s solution . anticoagulant . CATALOG NO QUANTITY
610 003 2 x 50 ml
Assay Principle Stability : 7 days at 2 – 8 C
o
610 004 2 x 100 ml
o
Haemoglobin is oxidized by potassium ferricyanide which is 4 days at 20 – 25 C
converted into stable cyanomethaemoglobin by potassium
cyanide. The absorbance of the cyanomethaemoglobin System Parameters
is monitored at 540 nm . Wavelength 540 nm (Hg 546 nm)
Optical path 1 cm
Assay type End-point
Reagents Direction Increase
Sample : Reagent Ratio 1 : 250
Reagent (R1) e.g . : Reagent volume 2.5 ml
Potassium ferricyanide 40 mmol/l Sample volume 10 ml
o
Potassium phosphate 50 mmol/l Temperature 20 – 25 C
Incubation time 5 minutes
Reagent (R2) Zero adjustment Against reagent blank
Potassium cyanide 77 mmol/l
Procedure
Harmful (Xn): R20/21/22: Harmful by inhalation, in contact with skin Pipette into test tubes
and if swallowed. S7: Keep container tightly closed. S28.1: After working solution 2.5 ml
contact with skin, wash immediately with plenty of water. Blood sample 10 µl
S45: In case of accident or if you feel unwell, seek medical advice
immediately. The amount of cyanide present in one bottle of reagent Mix well and rinse the blood pipette several times with the
o
is apreciably less than the minimum lethal dose for an adult. reagents , and incubate for 5 minuts at 20-25 C . Measure
However, hydrogen cyanide is liberated by acidification. Never allow absorbance of specimen (Aspecimen) against reagent blank .
reagent to come in contact with acid.
Calculation
For further information, refer to the Haemoglobin plus reagent Haemoglobin concentration (g/dL) = Aspecimen x 36.77
material safety data sheet.
Haemoglobin concentration (mmol/L)= Aspecimen x 22.83
Reagents also contain non-reactive stablizers and surfactants
44 45
Haemoglobin (Ready-To-Use) SYMBOLS IN PRODUCT LABELLING
Performance Characteristics References
Drabkin’s Solution EC REP Authorised Representative Use by/Expiration Date Precision

1. International committee for standardization in haematology.
Brit. J. Haemat., 1967:13 (Suppl.) 71.
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions Within run (Repeatiblity)
LOT Batch Code/Lot number for use 2. Van Kampen, E. J. and Zijlstra, W.G., Clin. Chem. Acta.,
REF:611 001 (1 x 250 ml) 100 test Level 1 Level 2
REF Catalogue Number Manufactured by 1961:6:538 – 544.
REF:611 002 (2 x 250 ml) 200 test n 20 20
Consult instructions for use (Xn) - Harmful
REF:611 003 (2 x 500 ml) 400 test 3. Tietz NW, Ed. Clinical guide to laboratory tests. 2ND ED.
Temperature Limitation Mean (mg/dL) 10 14 Philadelphia: WB Saunders; 1990:566.
CV% 2.3 1.3
Intended Use Reagent Preparation Storage and Stability

Spectrum Diagnostics haemoglobin reagent is intended for the in- Reagent is supplied ready to Use.
vitro quantitative, diagnostic determination of haemoglobin in human Run to run (Reproducibility) ORDERING INFORMATION
blood. Reagent is stable until expiration date stated on label when stored Level 1 Level 2 CATALOG NO QUANTITY
o
at 15 - 25 C.Once opened, the opened vial is stable for 6 months at n 20 20 611 001 1 x 250 ml
Background the specified temperature. 611 002 2 x 250 ml
Mean (mg/dL) 11.1 14.1
Haemoglobin (Hb) is the red pigmented protein located in the 611 003 2 x 500 ml
CV% 2.9 2.1
er-ythrocytes and consists of four subunits. Its main function is Specimen Collection and Preservation
the transport of oxygen and carbon dioxide in blood. In normal Anticoagulated venous or capillary blood . Blood may be anticoa-
humanadults, at least 96 % of the haemoglobin is HbA. HbA2 is gulated with EDTA, or fluoride. Blood can be taken directly from a Methods Comparison
usually about 2.5 – 3 % of total haemoglobin. Fetal hemoglobin (HbF) finger or heel puncture without use of anticoagulant. A comparison between Spectrum Diagnostics Haemoglobin reagent
predominates during fetal life and diminishes rapidly during the first and a commercial reagent of the same methodology was performed
o
year of postnatal life. In normal adults less than 1 % is HbF.Blood Stability : 7 days at 2 – 8 C on 40 human blood samples. A correlation (r) of 0.983 was obtained.
o
haemoglobin concentration may be diminished as a consequence of 4 days at 20 - 25 C
hemorrhage or hemolysis or as a result of impaired blood formation Expected values
in the bone marrow. System Parameters 1- 6 days 15.2 – 23.5 g/dL ( 9.4 – 14.6 mmol/L)
14 – 50 days 10.3 – 16.6 g/dL ( 6.4 – 10.3 mmol/L)
Method Wavelength 540 nm (Hg 546 nm)
2 - 10 months 10.0 – 12.9 g/dL ( 6.1 – 8.0 mmol/L)
colorimetric method using Drabkin’s solution. Optical path 1 cm
1 – 15 years 11.0 – 14.3 g/dL ( 6.8 – 8.8 mmol/L)
Assay Principle Direction Increase Adults Women 12.0 – 16.0 g/dL (7.5 – 9.9 mmol/L)
Haemoglobin is oxidized by potassium ferricyanide which is Sample : Reagent Ratio 1 : 250 Men 14.0 – 18.0 g/dL (8.7 – 11.2 mmol/L)
converted into stable cyanomethaemoglobin by potassium cyanide. e.g .: Reagent volume 2.5 ml
The absorbance of the cyanomethaemoglobin is monitored at 540 Sample volume 10 ml Spectrum Diagnostics does not interpret the results of a
o
nm. Temperature 20 – 25 C clinical laboratory procedure; interpretation of the results is
Incubation time 5 minutes considered the responsibility of qualified medical personnel.
Reagents Zero adjustment Against reagent blank
Reagent Procedure
Potassium ferricyanide 0.62 mmol/l Pipette into test tubes Waste Disposal
Potassium phosphate 1.04 mmol/l Reagent 2.5 ml This product is made to be used in professional laboratories.
Potassium cyanide 1.54 mmol/l Blood sample 10 µl Please consult local regulations for a correct waste disposal.
Surfactant < 0.1 % S56: dispose of this material and its container at hazardous or
Harmful (Xn): R20/21/22: Harmful by inhalation, in contact with skin Mix well and rinse the blood pipette several times with the reagents, special waste collection point.
o
and if swallowed. S7: Keep container tightly closed. S28.1: After and incubate for 5 minuts at 20-25 C. Measure absorbance of S57: use appropriate container to avoid environmental
contact with skin, wash immediately with plenty of water. specimen (Aspecimen) against reagent blank. contamination.
S45: In case of accident or if you feel unwell, seek medical advice S61: avoid release in environment. refer to special instructions/
immediately. The amount of cyanide present in one bottle of reagent Calculation safety data sheets.
is apreciably less than the minimum lethal dose for an adult. Haemoglobin concentration (g/dL) = Aspecimen x 36.77
However, hydrogen cyanide is liberated by acidification. Never allow
reagent to come in contact with acid. Haemoglobin concentration (mmol/L)= Aspecimen x 22.83
For further information, refer to the Haemoglobin reagent material Quality Control
safety data sheet. Normal & abnormal commerical control blood of known
46 47
Lactate – Liquizyme
o
Mix and incubate for 5 minutes at 37 C or 10 minutes at 15 – 25 Reference
SYMBOLS IN PRODUCT LABELLING o
C. Measure absorbance of specimen (Aspecimen) and standard
(Astandard) against reagent blank within 30 minutes. 1. Bailey EM, Domenico P,Cunha BA. Bacterial or viral meningitis?
REF: 274 001 (5 x 20 ml) 100 test Measuring lactate in CSF can help you know quickly. Meningitis.
IVD For in-vitro diagnostic use Use by/Expiration Date
1990;88:217-223.
LOT Batch Code/Lot number CAUTION. Consult instructions Calculation (Aspecimen)
Catalogue Number for use Lactate conc. (mg/dL) = x 10 2. Field M, Block JB, Levin R, Rall DP. Significance of blood lactate
REF
(Astandard)
Consult instructions for use Manufactured by elevations amoung patients with acute leukemia and other
Intended Use
Quality Control neoplastic proliferative disorders. Am J Med. 1996;40:528-547.
Spectrum Diagnostics liquizyme Lactate reagent is intended for the
in-vitro quantitative, diagnostic determination of lactate in human Reagent (R2) contains sodium azide which may react with copper 3. Klein TO. Nervensysteme. In:Greiling H, Gressner AM,eds.
Plasma and CSF on both automated and manual systems. or lead plumbing. Lehrbuch der Klinischen Chemie und Pathobiochemie.
Stuttgart:Schattauer; 1987:859-893.
Methods Comparison
Background Reagent Preparation A comparison between Spectrum Diagnostics Lactate reagent and a 4. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER,
Lactic acid, present in blood entirely as lactate is an intermediary
commercial reagent of the same methodology was performed on 20 eds. Tietz Fundamentals of Clinical Chemistry. 4 th ed.
product of carbohydrate metabolism and is derived mainly from Prepare working solution as following:
human sera. A correlation of 0.992 was obtained. Philadelphia:WB Saunders;1996:351-374.
muscle cells and erythrocytes. The blood lactate concentration is
affected by its production in muscle cells and erythrocytes and its REF:274 001:add 2 ml from R2 to one bottle of R1; mix gently. 5. Sacks DB. Carbohydrates in: Burtis CA, Ashwood ER, eds.
rate metabolism in the liver. During exercise, blood lactate can Sensitivity
Tietz Textbook of Clinical Chemistry. 2 nd ed. Philadelphia:WB
increase up to ten times of normal levels. Under normal conditions, When run as recommended, the minimum detection limit of the
Or prepare the working solution according to the number of tests Sander; 1994;928-1001.
the ratio between lactate and pyruvate is constant(10:1).The l i v e r assay is 0.3 mg/dL (0.033 mmol/L).
required by mixing 9 volumes of reagent 1 (R1) and 1 volume of
can normally metabolize more lactate than is produced. reagent 2 (R2), e.g. 900 ml R1 +100 ml R2. 6. Tietz NW, ed. Clinical Guide to laboratory tests. 3 rd ed.
In the case of decreased perfusion of the liver , however, removal Linearity Philadelphia: WB Saunders;1995:351-374.
of lactate by the liver may be significantly reduced.The amount The reaction is linear up to lactate concentration of 90 mg/dL
Reagent Storage and Stability
of lactate in cerebrospinal fluid normally parallels blood levels. (9.99 mmol/L), specimens showing higher concentration should
All reagents are stable until expiration date stated on label when
CSF lactate level is increased in bacterial meningitis, epilepsy, o be diluted 1+1 using physiological saline, reassayed and the result
stored refrigerated at 2 – 8 C. Working solution is stable for 3 ORDERING INFORMATION
and intracranial hemorrhage. CSF lactate level may be an aid to o o multiplied by two (2).
months at 2 – 8 C or 1 week at 15 – 25 C.
distinguish between bacterial from viral meningitis. CATALOG NO QUANTITY
Deterioration Interfering substance 274 001 5 x 20 ml
Method Plasma
The working reagent is normaly clear or pale pink. Do not use
Enzymatic colorimetric method (LOX / PAP)with lactate oxidase and liquizyme lactate reagent if it is turbid or if the absorbance is greater
4-aminoantipyrine. Haemolysis
than 0.1 at 546 nm.
Haemoglobin levels higher than 2.5 g/L (0.16 mmol/L) increase the
apparent lactate concentration significantly.
Assay Principle Specimen Collection and Preservation
Lactate is oxidized to pyruvate and hydrogen peroxide (H2O2) Plasma and CSF. Do not use serum specimens. Avoid icteric and
Icterus
by lactate oxidase (LOX). In the presence of peroxidase (POD), haemolytic specimens.The only acceptable anticoagulants are
Bilirubin levels higher than 4.0 mg/dL (68 mmol/L) decrease apparent
hydrogen peroxide reacts with 2,4,6-tribromo-3-hydroxybenzoic acid fluoride/heparin and iodoacetate/heparin. Collection of satisfactory
lactate concentration significantly.
(THB) and 4-aminoantipyrine (4-AAP) to form a red quinoneimine specimen for lactate analysis requires special procedures to prevent
dye. changes of lactate both while and after the specimen is drawn. The
Lipemia
patient should be fasting and at complete rest and exercise of the
Pyruvate No significant interference.
Lactate LOX arm or hand should be avoided before or during collection of the
+ +
H2O2 specimens. The collected blood should be cooled on ice immediately
O2
and separated from the cells within 15 minutes. Once the plasma is
Ascorbic acid
separated from the cells, lactate values are stable.
Physiological ascorbic acid concentrations do not interfere with the
2H2O + 4-AAP Use the CSF samples with addition of glycolysis inhibitor, e.g.sodium
POD quinoneimine dye o test. Ascorbic Acid levels higher than 5 mg/dL (284 mmol/l) decrease
+ + fluoride. Lactate in CSF is stable for 3 hours at 20 – 25 C, for 24
o o the apparent lactate concentration significantly.
THB 4H2O hours at 4 – 8 C, and for 2 months frozen at -20 C, stable in plasma
o o
for 2 hours at 20 – 25 C and 2 days at 4 – 8 C.
Expected Values
The color intensity of the formed red quinoneimine dye is directly System Parameters Plasma Venous 4.5 – 19.8 mg/dL 0.5 – 2.2 mmol/L
proportional to the lactate concentration. It is determined by mea-
Arterial 4.5 – 14.4 mg/dL 0.5 –1.6 mmol/L
suring the increase in absorbance at 546 nm. Wavelength 546 nm
Optical path 1 cm
CSF Adult 10 – 22 mg/dL 1.1 – 2.4 mmol/L
Reagents Assay type End-point
Direction Increase Neonates 10 – 60 mg/dL 1.1 – 6.7 mmol/L
Standard lactate (ST) 10 mg/dL Sample : Reagent Ratio 1 : 100
e.g.: Reagent volume 1 ml Spectrum Diagnostics does not interpret the results of a
Reagent 1 (R1 Buffer) Sample volume 10 ml clinical laboratory procedure; interpretation of the results is
o o
Tris buffer 100 mmol/L Temperature 37 C or 15 – 25 C considered the responsibility of qualified medical personnel.
2,4,6-tribromo-3-hydroxybenzoic acid 2.0 mmol/L Zero adjustment Reagent blank All indications of clinical significance are supported by
o
4-Amino antipyrine 0.8 mmol/L Incubation time 5 minutes at 37 C or literature references.
o
10 minutes at 15 – 25 C
Reagent 2 (R2 Enzyme) Reagent Blank Limits Low 0.00 AU
Lactate oxidase >20 U/L High 0.25 AU Analytical Range
Peroxidase >15 U/L Sensitivity 0.3 mg/dL (0.033 mmol/L) 0.3 – 90 mg/dL ( 0.033 – 9.99 mmol/L).
Sodium Azide 0.02 % Linearity 90 mg/dL (9.99 mmol/L)
Waste Disposal
For further information, refer to the Lactate reagent material safety Procedure This product is made to be used in professional laboratories.
data sheet. Please consult local regulations for a correct waste disposal.
Precautions and Warnings Working special waste collection point.
1.0 ml 1.0 ml 1.0 ml
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Reagent S57: use appropriate container to avoid environmental
immediately with plenty of soap and water. In case of severe injuries; contamination.
Standard ------ 10 µl ------
seek medical advice immediately. S61: avoid release in environment. refer to special instructions/
Sample ------ ------ 10 µl safety data sheets.
48 49
Micrototal Protein (MT-P) SYMBOLS IN PRODUCT LABELLING
Level 1 Level 2 1. Henry R.J., Cannon, D.C., Winkelman J.W., “Clinical
Pyrogallol - Red EC REP Authorised Representative
CAUTION. Consult instructions n 20 20 Chemistry,Principles and Techniques.” Harper & Row, 2nd
Ed.1974.
REF:282 001 (2 x 50 ml) 100 test LOT Batch Code/Lot number for use Mean (mg/dL) 39 109
REF:282 002 (4 x100ml) 400 test REF Catalogue Number Manufactured by 2. Watanabe N., Kamei, S., Oh Kubo A., and Tok uda K., Clin,
SD 0.79 1.36
REF:282 003 (2 x100ml) 200 test Consult instructions for use (Xn) - Harmful Chem.,1986., 32/8:1551.
Temperature Limitation CV% 2.7 1.9
Intended Use
Spectrum Diagnostics microprotein reagent is intended for the in- o o Methods Comparison
Stability (urine): 1 day at 15 – 25 C; 8 weeks at 4 – 8 C; A comparison between Spectrum Diagnostics Micrototal Protein
vitro quantitative, diagnostic determination of total protein in human 1 year at
o
-20 C
cerebrospinal fluid (CSF) and urine on both automated and manual reagent and a commercial reagent of the same methodology was ORDERING INFORMATION
systems. o o performed on 20 human Urine samples. A correlation of 0.975 was
Stability (CSF): 1 day at 15 – 25 C; 4 weeks at 4 – 8 C; obtained. CATALOG NO QUANTITY
o
6 months at -20 C
Background 282 001 2 x 50 ml
Protein level in spinal fluid may be increased in a variety of diseases Sensitivity 282 002 4 x 100 ml
System Parameters When run as recommended, the minimum detection limit of the
including tumors, meningitis, and polyneuritis. Most of the proteins 282 003 2 x 100 ml
Wavelength 600 nm (578 is an optional) assay is 6 mg/dL.
found in CSF originate from plasma; only 20% originate from Optical path 1 cm
intrathecal synthesis. The two main proteins found in human urine Assay type End-point
are albumin and uromucoid. Increased urinary proteins may be Linearity
Direction Increase
associated with a number of diseases, among them are destructive The reaction is linear up to microprotein concentration of 500 mg/
lesion of the kidney, primary and secondary nephropathies and also dL. Specimens showing higher concentration should be diluted 1+1
during pregnancy. using physiological saline and repeat the assay (result × 2).
Sample volume 20 ml
o
Temperature 15 – 25 C
Method Zero adjustment Reagent blank Interfering Substances
Colorimetric method (Pyrogallol–red molybdate complex). Incubation time
o
10 minutes ( 15– 25 C ) Avoid erythrocyte contamination. There is no significant interference
Sensitivity 6 mg/dL from the following substances present in urine up to the following
Assay Principle Linearity 500 mg/dL concentrations:
In acidic medium, protein in the specimen reacts with pyrogallol Uric Acid 85 mg/dL ( 5 mmol/ L)
red in the presence of molybdate ions to form a purple color complex. Procedure Oxalate 90 mg/dL (10 mmol/L)
This color complex absorbs maximally at 600 nm and the optical
density is directly proportional to protein concentration of the test Blank Standard Sample Phosphate 1.2 g /L (39 mmol/L)
sample. Reagent 1.0 ml 1.0 ml 1.0 ml Calcium 130 mg/dL ( 32 mmol/L)
Standard ------ 20 µl ------ Creatinine 6 g/L ( 53 mmol/L)
Reagents
Standard protein (ST) Sample ------ ------ 20 µl Ascorbic acid 10 mg/dL ( 568 mmol/L)
150 mg/dL
Mix, incubate for exactly 10 minutes at 15 - 25 C. Measure
o
Bilirubin 60 mg/dL (1.0 mmol/L)
Reagent (R) absorbance of specimen (Aspecimen) and standard(Astandard)
Succinate buffer 100 mmol/L against reagent blank within 10 minutes. Expected values
Sodium oxalate 4.0 mmol/L
Sodium molybdate 60 mmol/L Calculation Urine ( 24 hrs ) : 20 – 145 mg/day
Pyrogallol red 80 mmol/L (Aspecimen)
Harmful (Xn): R20/22: Harmful by inhalation and if swallowed. CSF or Urine protein conc. (mg/dL)= x 150 Urine ( random) : < 10 mg/dL
S24/25: Avoid contact with skin and eyes. (Astandard)
Concentration / 24hr urine: CSF : 15 – 45 mg/dL
For further information, refer to the Microprotein M-TP reagent
material safety data sheet. Measure the urine volume and calculate the concentration of M-TP Spectrum Diagnostics does not interpret the results of a
in 24hr urine as following: clinical laboratory procedure; interpretition of the results
Precautions and Warnings ml of urine is considered the responsibality of qualified medical
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Concentration of MTP /24hr urine = Concentration /dL x personnel. All indications of clinical significance are
100
immediately with plenty of soap and water. In case of severe injuries; supported by literature references.
seek medical advice immediately. Quality Control
Normal & abnormal control serum of known concentrations should
be analyzed with each run. Analytical Range
Spectrum Microprotein reagents are supplied ready-to-use and 6 – 500 mg/dL.
stable up to the expiry date labeled on the bottles,when stored at Performance Characterstics
o
2 – 8 C. Precision Waste Disposal
Deterioration Within run (Repeatiblity)
Please consult local regulations for a correct waste disposal.
Do not use the Spectrum microprotein reagents if turbid. Failure Level 1 Level 2 S56: dispose of this material and its container at hazardous or
to recover control values within the assigned range may be an special waste collection point.
indication of reagent deterioration. n 20 20
S57: use appropriate container to avoid environmental
Mean (mg/dL) 37 105 contamination.
Specimen Collection and preservation S61: avoid release in environment. refer to special instructions/
SD 0.74 1.27
Use Urine and CSF free from blood contamination. Urine:24 safety data sheets.
hour urine is the specimen of choice. Centrifuge urine specimen CV% 2.0 1.3
if turbidity is obvious. No special additives are required but keep
the specimen cool during collection. To avoid enhanced albumin
excretion, samples should not be collected after exertion or following
acute ingestion of a fluid load. CSF: Avoid blood contamination
since protein concentration in whole blood is 1000 times higher than
normal CSF. Centrifuge CSF specimen if turbidity is obvious.
50 51
PYRUVATE SYMBOLS IN PRODUCT LABELLING CALCULATION
(Quantitative Enzymatic UV–Test) EC REP Authorised Representative Temperature Limitation with Factor :
REF: 335 001 100 test Pyruvate (mg/dL) = DA x 6,37 ( at 340 nm )
R1 : 2 x 50 ml with Standard :
R2 : 1 x 5 ml
(DAsample)
R3 : 1 x 5 ml Pyruvate (mg/dL) = x 4.0
(DAstandard)
St : 1 x 20 ml
Expected values
0.3 – 0.7 mg/dL
Intended Use Reagent Storage and Stability Note: It is recommended for each laboratory to establish and
Spectrum Diagnostics liquizyme Pyruvate reagent is intended for the All reagents are stable until expiration date stated on label when maintain its own reference values. The given data are only an
o
in-vitro quantitative, diagnostic determination of pyruvate in human protected from light stored refrigerated at 2 - 8 C. indication.
blood.
Specimen Preparation QUALITY CONTROL
Method Pipet 2,0 mL of freshly drawn blood into a centrifugation tube For quality control use adequate control materials,
Enzymatic UV - Test. containing 4 mL of cold 0.6 m perchloric acid. Vortex for about 30 available from Spectrum Diagnostics.
seconds.
Assay Principle Keep the blood precipitate mixture for about 5 min in the cold Waste Disposal
In the presence of an excess of NADH pyruvate is converted to to assure complete protein precipitation. Centrifuge 10 min at This product is made to be used in professional laboratories.
lactate.The reduction of the absorbance = DA,at 340 nm, due to approximately 1500 x g. The protein free supernatant is ready for Please consult local regulations for a correct waste disposal.
the oxidation of NADH to NAD+ , is a measure of the amount of use. S56: dispose of this material and its container at hazardous or
pyruvate originally present : The Standard solution has to be diluted with perchloric acid, in the special waste collection point.
same ratio as the sample. S57: use appropriate container to avoid environmental
LDH
Pyruvate + NADH + H+ L-Lactate + NAD+ contamination.
System Parameters S61: avoid release in environment. refer to special instructions/
Reagents Wavelength 340 nm ( 334 - 365 ) safety data sheets.
Optical path 1 cm
Standard (St.) 4.0 mg/dl Assay type End point
Reagents: Spectrum Diagnostics does not interpret the results of a
o o
Temperature 30 C or 37 C clinical laboratory procedure; interpretation of the results is
R1 Buffer Reagent : Zero adjustment Against Water considered the responsibility of qualified medical personnel.
(Tris buffer, pH 7.20) 1.50 mol/L Linearity 400 mg/dl (61.2 mmol/l)
R2 Coenzyme Procedure
NADH 10.0 mmol/L Sample Standard
R1 Buffer Reagent 1 ml 1 ml
R3 Start Reagent ORDERING INFORMATION
Supernatant Sample 2 ml ------
LDH 1.50 kU/mL CATALOG NO QUANTITY
Diluted Standard ------ 2 ml
Mix and add 50 ml 50 ml 335 001 100 Test
Additional Reagent ( not provided with the kit )
R2 Coenzyme
Perchloric Acid 0.6m for deproteinization
Mix and incubate for approximately 5 min ,pour into uvette,measure
initial absorbance A1
Do not ingest or inhalate. In case of contact with eyes or skin; rinse R3 Start Reagent 50 ml 50 ml
immediately with plenty of soap and water. In case of severe injuries; mix, incubate for approximately 5 min and measure absorbance
A2
Reagent Preparation DA = A2 - A1 For Sample and Standard

Spectrum Pyruvate reagents are supplied ready-to-use.
52 53
Total bile acids (TBA) SYMBOLS IN PRODUCT LABELLING
Level 1 Level 2 1. Abbassi-Ghanavati M, Greer LG, cunningham FG.
Enzymatic recycling method EC REP Authorised Representative Temperature Limitation
n 20 20
IVD For in-vitro diagnostic use Use by/Expiration Date 2. Wallach, J.,Eighth ed. lippincott Williams & Wilkins
REF: 305 001 100 test LOT Batch Code/Lot number CAUTION. Consult instructions Mean (µmol/L) 85.16 228.18
3. H. Bergmeyer, K. Gawehn, and M. Grassl in Methods of
SD 3.25 6.69 Enzymatic Analysis (Bergmeyer H. U. ed) 2nd Volume I, 505-
CV% 3.82 2.93 507, Academic Press, Inc. New York, NY (1974).
4. Drummond, G. I. & Masanobu, Y. In: The enzymes (boyer, P.D.
(ed.), (3rd ed.), vol.4, pp. 337(1971).
Reagent Preparation, Storage and Stability Calibration
Intended Use Spectrum TBA reagents are supplied ready-to-use and stable up to
Total bile acids (TBA) diagnostic reagent is used for quantitative The assay requires the use of a total bile acids calibrator.
the expiry date labeled on the bottles at 2 – 8 oC.
in vitro determination of total bile acids in serum or plasma on Recalibration is recommended at anytime if one of the following
photometric systems. events occurs:
Deterioration * The Lot number of reagents changes.
Failure to recover control values within the assigned range may be ORDERING INFORMATION
Background * Preventative maintenance is performed or a critical
an indication of reagent deterioration. component is replaced. CATALOG NO QUANTITY
Liver: Total bile acids are metabolized in the liver and hence serve
as a marker for normal liver function. Total bile acids are increased * Control values have shifted or are out of range and a new
Specimen Collection and Preservation vial of control does not rectify the problem. 305 001 100 test
in patients with acute hepatitis, chronic hepatitis, liver sclerosis and
liver cancer, liver cancer, Cholestasis, Congenital and acquired The only acceptable anticoagulat is EDTA. Use preferably fresh
vascular shunts. serum. Methods Comparison
A comparison between Spectrum Diagnostics TBA reagent and a
Pregnancy: Obestetric cholestasis is the most common liver Stability: commercial reagent of the same methodology was performed on 20
disease seen during pregnancy and the test key diagnostics tool 1 week at 4 – 8 oC human sera. A correlation of 0.97 was obtained.
for investigating suspected cases. the main symptom is itching, but 2 months at -20 oC
tiredness , nausea and jaundice may also be reported. in addition Sensitivity
to the discomfort experience by the mother, high levels of bile acids System Parameters When run as recommended, the minimum detection limit of this
also increased risk to the baby and may lead to premature birth or Wavelength 405 nm assay 0.5 µmol/L.
greater risk of passing meconium in the womb. Optical path 1 cm
Assay type Fixed rate Linearity
Veterinary: In veterinary medicine the bile acids test is typically used Direction Increase The reaction is linear up to an albumin concentration of 180 µmol/L
to assess liver function in mammals such as cats, dogs, horses, pigs, Temperature 37 oC ; specimens showing higher concentration should be diluted 1+1
cows and sheep. The test is also of use in the investigation of avian Incubation time 5 minutes at 20–25oC with physiological saline and repeat the assay (result × 2).
liver disease, where elevated liver enzymes do not always correlate Zero adjustment Reagent Blank
with liver disease. In the most species a single, random sample is Sensitivity 0.5 µmol/L
assessed but a dual sampling protocol is typically used with cats Linearity 150 µmol/L
Serum, plasma
and dogs to investigate bile reabsorption. For the latter a fasting
blood sample is taken and then a second, post prandial sample Procedure Haemolysis
taken a few hours after being fed for comparison. Total bile acids are
Blank Calibrator Specimen A haemoglobin level of 150 mg/dL results in 13 % positive bias.
typically low in fasted state, whilst level will rise immediately after a
meal. When the liver has normal mass and blood flow the bile acid Reagent1(R1) 300 ml 300 ml 300 ml Icterus
levels will rapidly fall back to near fasting level as the liver removes
Calibrator ----- 5 ml ----- No significant interference up to a bilirubin level of 30 mg/dL.
them from the blood and returns them to the gall bladder.
Specimen ----- ----- 5 ml
Lipemia
Method Mix , incubate for 5 minutes at 37oC then add: No significant interference up to an intralipid level of 2 %.
Enzymatic recycling method
Reagent2(R2) 100 ml 100 ml 100 ml
Expected Values
Assay Principle Mix, and after 1 minute read the absorbance A1 of the Calibrator or 0 -10.4 µmol/L
In the presence of Thio-NAD, the enzyme 3-á hydroxysteroid specimen. After 3 minutes later, read absorbance A2 of calibrator It is recommended that each laboratory should establish its own
dehydrogenase (3-á HSD) converts bile acids to 3-keto steroids and or specimen. reference interval.
Thio-NADH. The reaction is reversible and 3-á HSD can convert
3-keto steroids and Thio-NADH to blie acids and Thio-NAD. In the Calculation Analytical Range
presence of excess NADH, the enzyme cycling occurs efficiently A2 – A1 = Aspecimen or Acalibrato or Ablank. 0.5 – 180 µmol/L.
and the rate of formation of Thio-NADH is determined by measuring
specific change of absorbance at 405nm. (ASpecimen - Ablank)
TBA(µmol/L) = x conc. of calibrator Waste Disposal
(ACalibrator - Ablank)
Reagents This product is made to be used in professional laboratories.
Quality Control Please consult local regulations for a correct waste disposal.
Reagent 1 (R1) Normal & abnormal commercial control serum of known S56: dispose of this material and its container at hazardous or
Buffer (pH 7.0) 65mmol/L concentrations should be analyzed with each run. special waste collection point.
Thio-NAD 1 g /L S57: use appropriate container to avoid environmental
Performance Characteristics contamination.
Reagent 2 (R2) S61: avoid release in environment. refer to special instructions/
Precision
Buffer (pH 7.0) 65 mmol/L safety data sheets.
Thio-NADH 6 g /L
Sodium Azide <0.1% Level 1 Level 2
3-a HSD >5000 U/L n 20 20
Precautions and Warnings Mean (µmol/L) 45.54 171.62

Do not ingest or inhalate. In case of contact with eyes or skin; rinse SD 0.35 0.65
CV% 0.76 0.38
54 55
Total protein SYMBOLS IN PRODUCT LABELLING
Level 1 Level 2 1. Cannon DC, Olitzky I, Inkpen JA :Proteins. In:Clinical
Biuret Reagent EC REP Authorised Representative
CAUTION. Consult instructions n 20 20 chemistry, principles and technics, 2 nd ed. RJ Henery, DC
Cannon, JW Winkelman, editors, Harper & Row, New York, pp
REF: 310 001 (2 x 100ml) 200 test LOT Batch Code/Lot number for use Mean (mg/dL) 5.7 7.32 407 – 421,1974.
REF: 310 002 (4 x 100ml) 400 test REF Catalogue Number Manufactured by
SD 0.19 0.21
REF: 310 003 (8 x 100ml) 800 test Consult instructions for use (C) - Corrosive 2. Gornall AG, Bardawill CJ, David MM: Determination of serum
REF: 310 004 (2 x 500ml) 1000 test Temperature Limitation CV% 2.53 2.4 protein by means of the biuret reagent. J Biol Chem 177:751,
REF: 310 005 (2 x 250ml) 500 test 1949 .
Methods Comparison
Deterioration 3. Kaplan A, Szalbo J :Clinical chemistry :Interpretation and
Intended Use A comparison between Spectrum Diagnostics Total Protein reagent
Do not use The total protein regents if precipitate forms. Failure to techniques, 2 nd ed. A Kaplan, J Szabo, editors, 1983, p 157.
Spectrum Diagnostics total protein reagent is intended for the in- and a commercial reagent of the same methodology was performed
recover control values within the assigned range may be an on 20 human sera. A correlation of 0.978 was obtained. 4. Schultze HE, Heremans JF:Molecular biology of human protein.
vitro quantitative, diagnostic determination of total protein in human indication of reagent deterioration.
serum on both automated and manual systems. Elsevier publishing company, Amsterdam, 1966.
Sensitivity
Specimen Collection and Preservation 5. Tietz NW : Fundamentals of Clinical Chemistry: 2 nd ed. NW
Background When run as recommended, the minimum detection limit of this
Tietz, editor, 1994, pp692 .
Plasma proteins are mainly synthesized in the liver and are involved assay is 1.0 g/dL.
Use serum or plasma ( EDTA or heparin ) for the test . Usually
in the maintenance of normal water distribution between tissues plasma results are higher due to fibrinogen . The serum or plasma
and blood, as well as acid-base balance. Due to some pathological Linearity
should be separated from the cells within 4 hours .
conditions, both total protein level and the ratio of different fractions o o The reaction is linear up to total protein concentration of 12 g/dL.
Stability : 1 day at 15 – 25 C ; 4 weeks at 4 – 8 C; ORDERING INFORMATION
may change independently of one another. Hyperproteinemia o Specimens showing higher concentration should be diluted 1+1
1 year at -20 C
may be detected during dehydration associated with diarrhea or using physiological saline and repeat the assay (result × 2). CATALOG NO QUANTITY
vomiting. The total protein levels also increase in multiple myeloma. System Parameters
Interfering Substances 310 001 2 x 100 ml
Hypoproteinemia may occur as a result of prolonged low protein diet Wavelength Hg 546 nm (530 – 570 nm)
Serum, plasma 310 002 4 x 100 ml
and in some pathological conditions such as nephrotic syndrome, Optical path 1 cm 310 003 8 x 100 ml
bleeding, sprue, and salt retention. Assay type End-point
Hemolysis 310 004 2 x 500 ml
Direction Increase 310 005 2 x 250 ml
Method No interference up to hemoglobin level of 7.5 g/L.
Coloremetric method (Biuret reagent). e.g .: Reagent volume 1 ml
Icterus
Sample volume 20 ml
Assay Principle o No significant interference up to a bilirubin level of 30 mg/dL.
Temperature 15 – 25 C
In alkaline medium the copper reacts with the peptide bonds of Zero adjustment Reagent blank
proteins to form the characteristic pink to purple biuret complex. o Lipemia
Incubation time 10 minutes at 15 – 25 C
Sodium potassium tartarate prevents copper hydroxide precipitation, No significant interference.
Sensitivity 1.0 g/dL
and potassium iodide prevents the autoreduction of copper. Linearity 12 g/dL
Drugs
2+ Alkaline pH Sera from patients receiving dextran may cause artificially high
Protein + Cu Cu – protein complex Procedure levels due to turbidity during color development. This positive bias
Blank Standard Sample can be minimized by centrifuging the reaction mixture before reading
The color intensity is directly proportional to the protein concentration. the absorbance.
It is determined by measuring the increase in the absorbance at Reagent (R) 1.0 ml 1.0 ml 1.0 ml
546nm. Standard ------ 20 µl ------ Expected Values
Reagents Sample ------ ------ 20 µl Adults 6.6 – 8.7 g/dL
Standard Total protein (ST) Children (> 1 year) 6.0 – 8.0 g/dL
6.0 g/dL Mix, Incubate for 10 minutes at room temp. Measure absorbance of
specimen (Aspecimen) and standard (Astandard) against reagent (< 1 year) 4.8 – 7.6 g/dL
Reagent (R) blank within 30 minutes. Newborns (< 4 weeks) 4.6 – 6.8 g/dL
Sodium hydroxide 750 mmol/L Prematures 3.4 – 5.0 g/dL
Copper sulfate 12.0 mmol/L Calculation
Sodium potassium tartarate 40.9 mmol/L (Aspecimen) Spectrum Diagnostics does not interpret the results of a
Potassium iodide 19.8 mmol/L Serum protein conc. (g/dL) = x6
(Astandard) clinical laboratory procedure;interpretation of the results is
(C)-Corrosive contains caustic materials.
Note: For turbid highly icteric sera, prepare a serum blank by All indications of clinical significance are supported by
R34 Causes burns.
adding 20 ml serum to 1 ml saline into a labeled test tube. Read literature references.
S26-45 In case of contact with eyes, rinse immediately with
plenty of water and seek medical advice in case of absorbance of serum blank at 540 nm vs water and subtract serum
accident or if you feel unwell, seek medical advice blank absorbance from test absorbance before calculating results. Analytical Range
immediately. 1.0 – 12 g/dL.
Quality Control
For further information, refer to the Total Protein reagent material Normal & abnormal commercial control serum of known Waste Disposal
safety data sheet. concentrations should be analyzed with each run. This product is made to be used in professional laboratories. Please
consult local regulations for a correct waste disposal.
Precautions and Warnings Performance Characteristics S56: dispose of this material and its container at hazardous or
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Precision special waste collection point.
immediately with plenty of soap and water. In case of severe injuries; Within run (Repeatiblity) S57: use appropriate container to avoid environmental
seek medical advice immediately. contamination.
Level 1 Level 2
Reagent Preparation, Storage and Stability n 20 20 safety data sheets.
Spectrum Total protein reagents are supplied ready-to-use and Mean (mg/dL) 5.2 7.23
stable up to the expiry date labeled on the bottles. The reagents
o
are stable at 15 – 25 C. Only the standard is needed to be kept SD 0.12 0.15
o
refrigerated at (2 - 8 C). CV% 2.47 2.2
56 57
Triglycerides – Liquizyme SYMBOLS IN PRODUCT LABELLING
Procedure Others
physiological ascorbic acid concentration dosn’ t interfere with
GPO-PAP (Single Reagent) EC REP Authorised Representative Temperature Limitation the test. Ascorbic acid levels higher than 114 mmol /l (2 mg /dL)
IVD For in-vitro diagnostic use Use by/Expiration Date Reagent 1.0 ml 1.0 ml 1.0 ml decrease the apparent triglycerides concentration significantly.
REF: 314 001 ( 2 x 25 ml) 50 test LOT Batch Code/Lot number CAUTION. Consult instructions Standard ------ 10 µl ------
REF: 314 002 ( 4 x 25 ml) 100 test REF Catalogue Number for use Expected Values
REF: 314 003 ( 3 x 40 ml) 120 test Sample ------ ------ 10 µl
REF: 314 004 (10 x 15 ml) 150 test o o
Females 35 -135 mg/dL (0.4 – 1.54 mmol/L)
REF: 314 005 ( 4 x 50 ml) 200 test Mix and incubate for 5 minutes at 37 C or 10 minutes at 15 – 25 C. Males 40 -160 mg/dL (0.45 – 1.82 mmo/L)
REF: 314 006 ( 4 x 100 ml) 400 test Reagents Measure absorbance of specimen (Aspecimen) and standard
REF: 314 007 ( 8 x 100 ml) 800 test Standard triglyceride (ST) (Astandard) against reagent blank within 30 minutes. The following limits are recommended for the recognition of the risk
REF: 314 008 ( 5 x 100 ml) 500 test 200 mg/dL 2.29 mmol/L factor hypertriglyceridemia:
REF: 314 009 ( 4 x 6 0 ml) 240 test Reagent (R) Calculation
Pipes Buffer pH 7.0 50 mmol/L (Aspecimen) Suspicious above 150 mg/dL (1.71 mmol/L)
4-chlorophenol 6.0 mmol/L Serum triglycerides conc. (mg/dL) = x 200 Elevated above 200 mg/dL (2.28 mmol/L)
Magnesium aspartate >0.5 mmol/L (Astandard)
Intended Use
Lipase >10 K U/L Quality Control
Spectrum Diagnostics liquizyme triglycerides reagent is intended Spectrum Diagnostics does not interpret the results of a
Peroxidase >2.0 KU/L Normal & abnormal commercial control serum of known
for the in-vitro quantitative, diagnostic determination of triglycerides clinical laboratory procedure;interpretation of the results is
4-Aminoantipyrine 1.0 mmol/L concentrations should be analyzed with each run.
in human serum on both automated and manual systems. considered the responsibility of qualified medical personnel.
Glycerol-3-phosphate oxidase >3.5 K U/L
Glycerol kinase >750 U/L Performance Characteristics
Background ATP 1.0 mmol/L
Triglycerides are the main lipids present in the human plasma; the Precision
Sodium Azide 8.0 mmol/L Within run (Repeatiblity)
others are the cholesterol, phospholipids and nonesterified fatty
Dynamic Range
acids. They are formed in the intestinal mucosa by the esterification Level 1 Level 2
For further information, refer to the Triglycerides reagent material 5 - 1000 mg/dL (0.057 - 11.45 mmol/L)
of glycerol and fatty acids. Triglycerides measurements are used in
safety data sheet.. n 20 20
the diagnosis and treatment of patients with diabetes mellitus, liver
obstruction, nephrosis and other diseases involving lipid metabolism. Waste Disposal
Precautions and Warnings Mean (mg/dL) 155.1 245.8
The measurement of serum triglycerides is important in the This product is made to be used in professional laboratories.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse SD 2.03 1.85 Please consult local regulations for a correct waste disposal.
diagnosis of hyperlipoproteinemia and in the prediction, detection
immediately with plenty of soap and water. In case of severe injuries; S56: dispose of this material and its container at hazardous or
and monitoring of atherosclerosis. CV% 1.31 0.75
seek medical advice immediately. special waste collection point.
Method Reagent (R) contains sodium azide which may react with copper or
contamination.
GPO-PAP-enzymatic colorimetric method. Level 1 Level 2
lead plumbing. S61: avoid release in environment. refer to special instructions/
n 20 20 safety data sheets.
Assay Principle Reagent Preparation, Storage and Stability
The series of the reaction involved in the assay system is as follows: Mean (mg/dL) 156 246.5
Spectrum triglyceride reagents are supplied ready-to-use and stable References
up to the expiry date labeled on the bottles when properly stored SD 2.2 1.9
1. Triglycerides are hemolyzed by lipoprotein lipase (LPL) to o
refrigerated at ( 2 – 8 C). Once opened, the opened vial is stable for 1. Bucolo G, David H : Quantitative determination of serum
glycerol CV% 1.4 0.87
3 months at the specified temperature. triglycerides by the use of the enzymes. Clin Chem 19 : 475,
LPL 1973
Triglycerides Glycerol + Fatty acids Methods Comparison
Deterioration 2. Chowdhury RF, Rodman H, Bleicher SJ : Glycerol like
The reagent is normally clear or pale pink. Do not use liquizyme A comparison between Spectrum Diagnostics Triglyceride reagent contamination of commerical blood sampling tubes. J Clin
2. Glycerol is then phosphorylated to glycerol-3-phosphate by and a commercial reagent of the same methodology was performed
triglyceride reagent if it is turbid or if the absorbance is greater than Pathol 12: 116, 1971
ATP in a reaction catalyzed by glycerol kinase (GK). on 20 human sera. A correlation of 0.967 was obtained.
0.2 at 546 nm. 3. MGowan MW, Artiss JD, Standbergh DR, Zak B. A peroxidase-
coupled method for colorimetric determination of serum
Glycerol Glycerol-3-phosphate Sensitivity
GK Specimen Collection and Preservation triglycerides. Clin Chem ;29:538-452 ;1983.
+ + When run as recommended, the minimum detection limit of the
Patients should be fasting for 10 to 14 hours before blood is drawn. 4. Stein EA; Lipids , lipoproteins, and apolipoproteins. In : NW Tietz
ATP ADP assay is 5 mg/dL (0.057 mmol/L).
Samples must be drawn in a soap and glycerol free collection device. , ed. Fundamentals of clinical chemistry, 3 rd ed. Philadelphia :
Recommended anticoagulats are EDTA or heparin at levels of 1mg WB Saunders; 448 ; 1987.
3. The oxidation of glycerol-3-phosphate is catalyzed by glycerol
and 0.2 mg/dl whole blood, respectively. Linearity 5. Tietz NW, Boden T, Stepleton JD : An improved method for the
phosphate oxidase (GPO) to form dihydroxyacetone phosphate
The reaction is linear up to triglycerides concentration of 1000 mg/ determination of lipase in serum. Am J Clin Pathol 31: 148,
and hydrogen peroxiode (H2O2).
Triglycerides in serum samples remain stable for 7 days at 4 C, for
o dL; specimens showing higher concentration should be diluted 1+1 1959
o o
3 months at -20 C, and for years at -70 C. using physiological saline and repeat the assay (result × 2). 6. Young DS et al, Clin Chem. 21 ; 1975
Glycerol-3-phosphate GPO Dihydroxyacetone phosphate
+ +
O2 H2O2 System Parameters Interfering Substances ORDERING INFORMATION
Wavelength Hg 546 nm (500 – 550 nm) Serum, plasma
Optical path 1 cm CATALOG NO QUANTITY
4. In the presence of peroxidase (POD), hydrogen peroxide effects
Assay type End-point Haemolysis 2 x 25 ml
the oxidative coupling of 4-chlorophenol and 4-aminoantipyrine 314 001
Direction Increase No significant interference up to a haemoglobin level of 6.0 g/L 4 x 25 ml
(4AAP) to form a red color quinoneimine dye which is measured 314 002
Sample : Reagent Ratio 1 : 100 (0.36 mmol/L). 3 x 40 ml
at 546 nm. 314 003
e.g.: Reagent volume 1 ml 10 x 15 ml
Icterus 314 004
2 H2O2+4-APP Quinoneimine dye Sample volume 10 µl 4 x 50 ml
POD o o Bilirubin levels higher than 171 mmol/L (10 mg/dL) decrease the 314 005
+ + Temperature 15 – 25 C or 37 C 4 x 100 ml
apparent triglycerides concentration significantly. 314 006
4-chlorophenol 4H2O Zero adjustment Reagent blank 8 x 100 ml
o 314 007
Incubation time 10 minutes at 15 – 25 C or 5 x 100 ml
o Drugs 314 008
5 minutes at 37 C 4 x 60 ml
Of the drugs tested in-vitro, methyldopa and levodopa cause 314 009
High 0.2 AU artificially low triglyceride values at the tested drug Level.
Sensitivity 5 mg/dL (0.057 mmol/L)
58 59
Urea/BUN - Liquizyme
o
Mix and incubate for 5 minutes at 37 C or 10 minutes at suppressed by amines, thiols, steroids and ascorbic acid.
SYMBOLS IN PRODUCT LABELLING 20-25 C. Measure absorbance of specimen (Aspecimen) and
o
(Modified Urease-Berthlot Method) EC REP Authorised Representative Use by/Expiration Date standard (Astandard) against reagent blank. Expected Values
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions
REF: 318 001 100 test REF: 318 002 200 test LOT Batch Code/Lot number for use Calculation Urea(Serum)
REF Catalogue Number Manufactured by (Aspecimen) Adults ≤ 65 years : 15 – 50 mg/dL (2.5-8.33 mmol/L)
R1 Buffer 1 x 100 ml R1 Buffer 2 x 100 ml Consult instructions for use (Xi) - Irritant Serum urea concentration (mg/dl) = A xn Adults ≥ 65 years : ≤ 70 mg/dL (≤11.66 mmol/L)
R2 Urease 1 x 6 ml R2 Urease 2 x 6 ml ( standard)
R3 Alkaline reagent 1 x 20 ml R3 Alkaline reagent 1 x 40 ml where n = 50.0 mg/dl (8.33 mmol/l) BUN(Serum)
Adults ≤ 65 years : 7 – 23.5 mg/dL
REF: 318 003 500 test REF: 318 004 1000 test NB: For megalabs having high numbers of patient specimens,working Urine urea concentration is determined by multiplying the result by Adults ≥ 65 years : 7 – 32.9 mg/dL
buffer reagent can be prepared .( Stability 1 week ) the dilution factor (50). Children : 5 – 18 mg/dL
R1 Buffer 5 x 100 ml R1 Buffer 4 x 250 ml REF:318 001: add 5 ml from R2 to one bottle of R1; mix gently.
R2 Urease 2 x 15 ml R2 Urease 51 ml REF:318 002: add 5 ml from R2 to one bottle of R1; mix gently. Urea Nitrogen: To convert the result from urea to urea nitrogen Urine (24) hours
R3 Alkaline reagent 2 x 50 ml R3 Alkaline reagent 1 x 210 ml REF:318 003: add 5 ml from R2 to one bottle of R1; mix gently. multiply the result by 0.467. Urea : 20 – 35 g/24hrs (330-580 mmol/24hrs)
REF:318 004:add 12.5 ml from R2 to one bottle of R1; mix gently. BUN : 9.3 – 16.4 g/24hrs
Intended Use
Spectrum Diagnostics colorimetric urea reagent is intended for Quality Control
the in-vitro quantitative, diagnostic determination of urea in human Deterioration Normal & abnormal control serum of known concentrations should Spectrum Diagnostics does not interpret the results of a
serum on both automated and manual systems. Do not use the reagent if it is turbid. Failure to recover control be analyzed with each run. clinical laboratory procedure; interpretation of the results is
values within the assigned range may be an indication of reagent
deterioration. Performance Characteristics
Background All indications of clinical significance are supported by
Urea is the major end product of protein nitrogen metabolism. It Precision literature references.
is synthesized by the urea cycle in the liver and excreted through Specimen Collection and Preservation Within run (Repeatiblity)
the kidneys. The circulating levels of urea depend upon protein Serum
No special preparation of the patient is required. Use nonhaemolyzed Level 1 Level 2 Analytical Range
intake, protein catabolism and kidney function. Elevated urea levels
serum or plasma only.The only acceptable anticoagulants are n 20 20 0.6 – 200 mg/dL (0.1 - 33.3 mmol/L).
can occur due to renal impairment or in some diseases such as
diabetes, infection, congestive heart failure and during different liver heprin, EDTA and fluoride. Do not use ammonium heparin plasma.
o o Mean (mg/dL) 60 144 Waste Disposal
diseases. Determination of blood urea nitrogen is the most widely Stability: 7 days at 15 –25 C ; 7 days at 2 – 8 C;
o
1 year at -20 C SD 1.87 2.1 This product is made to be used in professional laboratories.
used screening test for renal function together with serum creatinine.
Urine Please consult local regulations for a correct waste disposal.
CV% 3.12 1.46 S56: dispose of this material and its container at hazardous or
Urine samples are prediluted 1 : 50 with ammonium free water prior
Method special waste collection point.
to assay.
Urease-colorimetric method. Run to run (Reproducibility)
o
Stability: 2 days at 15 –25 C ; 7 days at 2 – 8 C;
o
1 month at -20 C
o
Level 1 Level 2 contamination.
Assay Principle S61: avoid release in environment. refer to special instructions/
The reaction involved in the assay system is as follows: n 20 20
System Parameters safety data sheets.
Urea is hydrolyzed in the presence of water and urease to produce Mean (mg/dL) 62 146
Wavelength 578 nm (578-623 nm)
ammonia and carbon dioxide.
Optical path 1 cm SD 1.92 2.5 References
Urease Assay type End-point 1. Batton, C. J & crouch, S.R : Anal. Chem., 1977,49:464-469.
Urea + H2O 2NH3 + CO2 CV% 3.25 1.65
Direction increase 2. Shephard MD, Mezzachi RD : Clin Biochem Revs, 4:61-7,
o
temperature 15-25 C or 37 C
o
1983.
The free ammonia in an alkaline pH and in the presence of indicator
Zero adjustment Against Reagent blank 3. Tietz NW, ED. Clinical guide to Laboratory tests. 2ND ED.
forms coloured complex proportional to the urea concentration in Methods Comparison
Reagent Blank Limits Low 0.02 AU Philadelphia: WB Saunders; 1990:566.
the specimen. A comparison between Spectrum Diagnostics Urea/BUN reagent
High 0.2 AU 4. Tiffany to, jansen JM, Burtis CA,Overton JB, Scott CD.
Sensitivity 0.6 md/dL (0.1 mmol/l) and a commercial reagent of the same methodology was performed Enzymatic Kinetic Rate and end Point analyses of
Reagents Linearity 200 mg/dL (33.3 mmol/l) on 20 human sera. A correlation of 0.97 was obtained. Substrate, By USE of A Gemsaec fast analyzer. Clin Chem.
Standard urea (ST) Aqueous primary standard
50 mg/dL 8.33 mmol/l Sensitivity
Procedure 1 ORDERING INFORMATION
When run as recommended, the minimum detection limit of the
Reagent 1 (R1 Buffer) Blank Standard Specimen assay is 0.6 mg/dL. CATALOG NO QUANTITY
Phosphate buffer pH 8.0 100 mmol/l
R1(Buffer) 1.0 ml 1.0 ml 1.0 ml 318 001 100 Test
Sodium salicylate 80 mmol/l Linearity
Sodium nitroprusside 6.0 mmol/l one drop one drop one drop 318 002 200 Test
R2(Enzyme) The reaction is linear up to a urea concentration of (200 mg/dl) 318 003 500 Test
EDTA 30.0 mmol/l (50 ml) (50 ml) (50 ml)
33.3 mmol/L. Specimens showing higher concentrations should 318 004 1000 Test
Standard ------ 10 ml ------ be diluted 1+2 with physiological saline and repeat the assay
Reagent 2 (R2 Enzyme) (result×3).
Urease >6000 U/l Sample ------ ------ 10 ml
o
Mix and incubate for at least 3 minutes at 37 C or 5 minutes at Interfering Substances
Reagent 3 (R3 Alkaline Reagent) o
20-25 C Serum, plasma
Sodium hydroxide 400 mmol/l
Sodium hypochlorite 20.0 mmol/l R3(Alk.Reagent) 200 ml 200 ml 200 ml
Haemolysis
Irritant (xi) R36/38: Irritating to eyes and skin. S26: In case of contact o
Mix and incubate for 5 minutes at 37 C or 10 minutes at Erythrocyte contamination doesn’t elevate results.
with eyes, rinse immediately with plenty of water and seek medical 20-25 C Measure absorbance of specimen (Aspecimen) and
o
advice. S37/39: Wear suitable gloves and eye/face protection. standard (Astandard) against reagent blank. Icterus
Procedure 2 (Using working solution) No significant interference.
For further information, refer to the Urea/BUN reagent material
safety data sheet. Blank Standard Specimen
Lipemia
Working solution 1.0 ml 1.0 ml 1.0 ml Lipemic specimens interfere with the method of Berthlot.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Standard ------ 10 ml ------
Anticoagulants
immediately with plenty of soap and water. In case of severe injuries; Sample ------ ------ 10 ml Ammonium heparin should not be used.
seek medical advice immediately. o
Mix and incubate for at least 3 minutes at 37 C or 5 minutes at
o
20 -25 C Others
Reagent Preparation, Storage and Stability Ammonium ions should be avoided since it may cause erroneously
Spectrum coloremetric urea reagents are supplied ready-to-use and R3(Alk.Reagent) 200 ml 200 ml 200 ml elevated results. Color development in the Berthlot reaction is
o
stable up to the expiry date labeled on the bottles (2 – 8 C).
60 61
Urea/BUN - LS SYMBOLS IN PRODUCT LABELLING
Quality Control
(Modified Urease- EC REP Authorised Representative
! CAUTION. Consult instructions
be analyzed with each run.
Berthlot Method) LOT Batch Code/Lot number for use Performance Characteristics literature references.
REF Catalogue Number Manufactured by Precision
Consult instructions for use (Xi) - Irritant Within run (Repeatiblity)
Temperature Limitation Analytical Range
REF: 321 001 90 Test REF: 321 002 180 Test Level 1 Level 2
0.6 – 200 mg/dL (0.1 - 33.3 mmol/L).
R1 Buffer 1 x 90 ml R1 Buffer 2 x 90 ml n 20 20
R2 Urease 1 x 1.5 ml R2 Urease 2 x 1.5 ml Mean (mg/dL) 60 144 Waste Disposal
R3 Alkaline reagent 1 x 20 ml R3 Alkaline reagent 1 x 40 ml This product is made to be used in professional laboratories.
SD 1.87 2.1 Please consult local regulations for a correct waste disposal.
REF: 321 003 270 Test REF: 321 004 400 Test
Reagent Preparation , Storage and Stability CV% 3.12 1.46 S56: dispose of this material and its container at hazardous or
R1 Buffer 3 x 90 ml R1 Buffer 4 x 100 ml To prepare the working solution add the content of one vial of special waste collection point.
R2 Urease 3 x 1.5 ml R2 Urease 4 x 1.7 ml urease (R2) to one bottle of buffer reagent (R1). S57: use appropriate container to avoid environmental
R3 Alkaline reagent 1 x 65 ml R3 Alkaline reagent 2 x 40 ml o
Stability : 1 Month at 2-8 C. contamination.
Level 1 Level 2 S61: avoid release in environment. refer to special instructions/
Intended Use Deterioration n 20 20 safety data sheets.
Spectrum Diagnostics colorimetric urea reagent is intended for Do not use the reagent if it is turbid. Failure to recover control
the in-vitro quantitative, diagnostic determination of urea in human Mean (mg/dL) 62 146
values within the assigned range may be an indication of reagent References
serum on both automated and manual systems. deterioration. SD 1.92 2.5
CV% 3.25 1.65 1. Batton, C. J & crouch, S.R : Anal. Chem., 1977,49:464-469.
Background Specimen Collection and Preservation 2. Shephard MD, Mezzachi RD : Clin Biochem Revs, 4:61-7,
Urea is the major end product of protein nitrogen metabolism. It Serum 1983.
is synthesized by the urea cycle in the liver and excreted through Methods Comparison
No special preparation of the patient is required. Use nonhaemolyzed A comparison between Spectrum Diagnostics Urea/BUN reagent 3. Tietz NW, ED. Clinical guide to Laboratory tests. 2ND ED.
the kidneys. The circulating levels of urea depend upon protein serum or plasma only.The only acceptable anticoagulants are Philadelphia: WB Saunders; 1990:566.
intake, protein catabolism and kidney function. Elevated urea levels and a commercial reagent of the same methodology was performed
heprin, EDTA and fluoride. Do not use ammonium heparin plasma. on 20 human sera. A correlation of 0.97 was obtained. 4. Tiffany to, jansen JM, Burtis CA,Overton JB, Scott CD.
can occur due to renal impairment or in some diseases such as o
o
Enzymatic Kinetic Rate and end Point analyses of Substrate,
diabetes, infection, congestive heart failure and during different liver 1 year at -20 C
o By USE of A Gemsaec fast analyzer. Clin Chem.
diseases. Determination of blood urea nitrogen is the most widely Sensitivity
Urine When run as recommended, the minimum detection limit of the
used screening test for renal function together with serum creatinine. Urine samples are prediluted 1 : 50 with ammonium free water prior assay is 0.6 mg/dL.
to assay.
Method o
o
Linearity ORDERING INFORMATION
Urease-colorimetric method. 1 month at -20 C
o
The reaction is linear up to a urea concentration of (200 mg/dl) CATALOG NO QUANTITY
Assay Principle 33.3 mmol/L. Specimens showing higher concentrations should
System Parameters be diluted 1+2 with physiological saline and repeat the assay 321 001 1 x 90 ml
The reaction involved in the assay system is as follows: Wavelength 578 nm (578-623 nm) (result×3). 321 002 2 x 90 ml
Urea is hydrolyzed in the presence of water and urease to produce Optical path 1 cm
ammonia and carbon dioxide. 321 003 3 x 90 ml
Urease Direction increase
Urea + H2O 2NH3 + CO2 o o Serum, plasma
temperature 15-25 C or 37 C
Zero adjustment Against Reagent blank
The free ammonia in an alkaline pH and in the presence of indicator Haemolysis
forms coloured complex proportional to the urea concentration in Erythrocyte contamination doesn’t elevate results.
High 0.2 AU
the specimen. Sensitivity 0.6 md/dL (0.1 mmol/l)
Icterus
Linearity 200 mg/dL (33.3 mmol/l)
Reagents No significant interference.

Standard urea (ST) Aqueous primary standard Procedure Lipemia
50 mg/dL 8.33 mmol/l
Blank Standard Specimen Lipemic specimens interfere with the method of Berthlot.
Reagent 1 (R1 Buffer) Working solution 1.0 ml 1.0 ml 1.0 ml
Anticoagulants
Phosphate buffer pH 8.0 100 mmol/l Standard ------ 10 ml ------ Ammonium heparin should not be used.
Sodium salicylate 80 mmol/l
Sodium nitroprusside 6.0 mmol/l Sample ------ ------ 10 ml
Others
EDTA 30.0 mmol/l Mix and incubate for at least 3 minutes at 37 oC or 5 minutes Ammonium ions should be avoided since it may cause erroneously
o
at 20-25 C . elevated results. Color development in the Berthlot reaction is
Reagent 2 (R2 Enzyme) suppressed by amines, thiols, steroids and ascorbic acid.
R3(Alk) 200 µl 200 µl 200 µl
Urease >350000 U/l
o
Mix and incubate for 5 minutes at 37 C or 10 minutes at 20-25 C
o
Expected Values
Reagent 3 (R3 Alkaline Reagent)
Sodium hydroxide 400 mmol/l Measure absorbance of specimen (Aspecimen) and standard
Urea(Serum)
Sodium hypochlorite 20.0 mmol/l (Astandard) against reagent blank .. Adults ≤ 65 years : 15 – 50 mg/dL (2.5-8.33 mmol/L)
Irritant (xi) R36/38: Irritating to eyes and skin. S26: In case of contact Adults ≥ 65 years : ≤ 70 mg/dL (≤11.66 mmol/L)
with eyes, rinse immediately with plenty of water and seek medical Calculation (Aspecimen)
advice. S37/39: Wear suitable gloves and eye/face protection. Serum urea concentration (mg/dl) = xn BUN(Serum)
(Astandard) Adults ≤ 65 years : 7 – 23.5 mg/dL
For further information, refer to the Urea/BUN reagent material where n = 50.0 mg/dl (8.33 mmol/l) Adults ≥ 65 years : 7 – 32.9 mg/dL
safety data sheet. Children : 5 – 18 mg/dL
Urine urea concentration is determined by multiplying the result by
Precautions and Warnings the dilution factor (50). Urine (24) hours
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Urea : 20 – 35 g/24hrs (330-580 mmol/24hrs)
immediately with plenty of soap and water. In case of severe injuries; Urea Nitrogen: To convert the result from urea to urea nitrogen BUN : 9.3 – 16.4 g/24hrs
seek medical advice immediately. multiply the result by 0.467.
62 63
Urea/BUN – Liquizyme (UV) SYMBOLS IN PRODUCT LABELLING
Calculation Expected Values
Urea (Serum)
EC REP Authorised Representative Temperature Limitation D A specimen = A1 specimen – A2 specimen Adults <65 years : 15-50 mg/dL (2.5-8.33 mmol/L)
REF: 319 001 (3 x 50 ml) 150 test
IVD For in-vitro diagnostic use Use by/Expiration Date D A standard = A1 standard – A2 standard Adults >65 years : < 70 mg/dL (<11.66 mmol/L)
REF: 319 002 (3 x 90 ml) 270 test
REF: 319 003 (4 x 100 ml) 400 test LOT Batch Code/Lot number CAUTION. Consult instructions (D Aspecimen)
Catalogue Number for use Serum urea concentration (mg/dl) = xn BUN (Serum)
REF: 319 004 (4 x 50 ml) 200 test REF
(D Astandard) Adults <65 years : 7-23.5 mg/dL
where n = 107 mg/dL Adults >65 years : 7-32.9 mg/dL
Intended Use Children : 5-18 mg/dL
Spectrum Diagnostics liquizyme urea reagent is intended for the in- Or prepare the working solution according to the number of tests Urine urea concentration is determined by multiplying the result
vitro quantitative, diagnostic determination of urea in human serum required by mixing 9 volumes of reagent 1 (R1) and 1volume of by the dilution factor (50). Urine (24) hours
or urine on both automated and manual applications. reagent 2 (R2) eg. 900 ml R1 +100 ml R2. Urea : 20-35 g/24hrs (330-580 mmol/24hrs)
Quality Control BUN : 9.3-16.4 g/24hrs
Background Precautions and Warnings Normal & abnormal commercial control serum of known
Urea is the major product of protein nitrogen metabolism. It is Do not ingest or inhalate. In case of contact with eyes or skin; rinse concentrations should be analyzed with each run.
synthesized by the urea cycle in the liver and excreted through immediately with plenty of soap and water. In case of severe injuries; Spectrum Diagnostics does not interpret the results of a
the kidneys. The circulating levels of urea depend upon protein seek medical advice immediately. clinical laboratory procedure;interpretation of the results is
Performance Characteristics considered the responsibility of qualified medical personnel.
intake, protein catabolism and kidney function. Elevated urea levels
can occur due to renal impairment or in some diseases such as Precision All indications of clinical significance are supported by
Both reagents (R1) and (R2) contain sodium azide which may react
diabetes, infection, congestive heart failure and during different liver Within run (Repeatiblity) literature references.
with copper or lead plumbing.
diseases. Determination of blood urea nitrogen is the most widely Level 1 Level 2
used screening test for renal function together with serum creatinine. Reagent Storage and Stability n 20 20 Analytical Range
All reagents are stable until expiration date stated on label when 0.9 – 200 mg/dL (0.15 - 49.8 mmol/L).
Method o
stored refrigerated at 2 – 8 C. Mean (mg/dL) 45 150
urease-UV fixed rate (enzymatic method). o
Working solution is stable for 1 month at 2 – 8 C or 8 days at SD 0.7 2.7 Waste Disposal
o
15 – 25 C. This product is made to be used in professional laboratories. Please
CV% 1.5 1.95
Assay Principle consult local regulations for a correct waste disposal.
The series of reactions involved in the assay are as follows : Deterioration S56: dispose of this material and its container at hazardous or
Do not use liquizyme BUN reagent if it is turbid or if the absorbance special waste collection point.
Run to run (Reproducibility) S57: use appropriate container to avoid environmental
1. Urea is hydrolyzed in the presence of water and urease to of the working reagent is less than 1.0 at 340 nm. Failure to recover
produce ammonia and carbon dioxide. control values within the assigned range may be an indication of Level 1 Level 2 contamination.
reagent deterioration. S61: avoid release in environment. refer to special instructions/
n 20 20
safety data sheets.
Urease
Urea + H2O 2NH3 + CO2 Specimen Collection and Preservation Mean (mg/dL) 47 153
No special preparation of the patient is required. Use nonhemolyzed SD 0.82 2.81 Referances
2. In the presence of glutamate dehydrogenase (GLDH) and serum or plasma only.The only acceptable anticoagulants are 1. Batton, C. J & Crouch, S.R: Anal. Chem. , 1977, 49:464-469 .
reduced nicotinamide adenine dinucleotide (NADH),the CV% 1.63 2.15 2. Shephard MD, Mezzachi RD: Clin Biochem Revs, 4:61-7, 1983.
heprin, EDTA and fluoride. Do not use ammonium heparin plasma.
ammonia combines with a-ketoglutarate (a-KG) to produce o
o
3. Tiffany TO, jansen JM, Burtis CA,Overtion JB, SCOTT CD.
L-glutamate. 1 year
o
at -20 C Enzymatic kinetic rate and end point analyses of substrate, by
Urine samples are prediluted 1 : 50 with ammonium free water prior Methods Comparison use of a gemsaec fast analyzer. Clin Chem. 1972;18:829-840.
2NH4 + 2a-KG 2 L-Glutamate to assay. A comparison between Spectrum Diagnostics Urea (UV) reagent 4. Tietz NW, Ed.Clinical guide to laboratory tests. 2ND.
GLDH and a commercial reagent of the same methodology was performed
+ + o
Stability: 2 days at 15 – 25 C ; 7 days at 2 – 8 C;
o
Philadelphia: WB Saunders;1990:566.
2 NADH 2 NAD+ + H2O 1 month at -20 C
o on 20 human sera. A correlation of 0.992 was obtained.
The rate decrease in the NADH concentration is directly proportional System Parameters Sensitivity
to the urea concentration in the specimen. It is determined by Wavelength 340 nm When run as recommended, the minimum detection limit of the CATALOG NO QUANTITY
measuring the absorbance at 340 nm. Optical path 1 cm assay is 0.9 mg/dL. 319 001 3 x 50 ml
Assay type Fixed Rate 319 002 3 x 90 ml
Reagents Direction Decrease Linearity 319 003 34 x 100 ml
Standard urea (ST) Sample : Reagent Ratio 1 : 100 The reaction is linear up to a urea concentration of 200 mg/dL. 319 004 4 x 50 ml
BUN 50 mg/dL e.g.: Reagent volume 1 ml Specimens showing higher concentration should be diluted 1+2
Urea 107 mg/dL Sample volume 10 ml with physiological saline and repeat the assay (result × 3).
First read time 30 seconds
Reagent 1 (R1 Buffer) Delay time 60 seconds Interfering Substances
Tris Buffer ( pH 8.5) 50 mmol/L Last read time 90 seconds Serum, plasma
a-Ketoglutarate 10 mmol/L Temperature 37 C
o
GLDH 8.0 K U/L Zero adjustment Against Air Haemolysis

Urease 5.0 K U/L Reagent Blank Limits Low 1.00 AU Erythrocyte contamination doesn’t elevate results.Haemolytic
Sodium azide 8.0 mmol/L High 2.0 AU specimens may cause high absorbance flagging.
Sensitivity 0.9 mg/dL (0.15 mmol/L)
Reagent 2 (R2 Starter) Linearity 300 mg/dL (49.8 mmol/L) Icterus
NADH >0.20 mmol/L No significant interference.
Sodium azide 8 mmol/L Procedure
Lipemia
For further information, refer to the Urea/Bun reagent material safety Standard Specimen Lipemic specimens may cause high absorbance flagging
data sheet. Working 1 ml 1 ml Diluted sample treatment may be recommended.
solution
Reagent Preparation Anticoagulants
Prepare working solution as following: Standard 10 µl ------ Ammonium heparin should not be used.
REF: 319 001 : add 5 ml from R2 to one bottle of R1; mix gently Specimen ------ 10 µl
REF: 319 002 : add one bottle from R2 to one bottle of R1 ; mix Others
gently Mix, and after 30 seconds read the absorbance A1 of the standard Ammonium ions should be avoided since it may cause
REF: 319 003 : add one bottle from R2 to one bottle of R1 ;mix or specimen. Exactly 1 minute later, read the absorbance A2 of erroneously elevated results.
gently standard or specimen.
REF: 319 004 : add 5 ml from R2 to one bottle of R1; mix gently
64 65
Urea/BUN – (UV) SYMBOLS IN PRODUCT LABELLING
Calculation
D A specimen = A1 specimen – A2 specimen
Expected Values
Ultimate Single Reagent EC REP Authorised Representative Temperature Limitation D A standard = A1 standard – A2 standard Urea (Serum)
IVD For in-vitro diagnostic use Use by/Expiration Date DAspecimen Adults <65 years : 15-50 mg/dL (2.5-8.33 mmol/L)
REF: 320 001 (2 x 20 ml) 40 test LOT Batch Code/Lot number CAUTION. Consult instructions Serum urea concentration (mg/dl) = xn Adults >65 years : < 70 mg/dL (<11.66 mmol/L)
DAstandard
REF: 320 002 (6 x 20 ml) 120 test REF Catalogue Number for use
where n = 107.0 mg/dL BUN (Serum)
REF: 320 004 (2 x 100 ml) 200 test Adults <65 years : 7-23.5 mg/dL
Urine urea concentration is determined by multiplying the result Adults >65 years : 7-32.9 mg/dL
by the dilution factor (50). Children : 5-18 mg/dL
Intended Use Precautions and Warnings
Spectrum Diagnostics Ultimate urea reagent is intended for the in- Do not ingest or inhalate. In case of contact with eyes or skin; rinse Quality Control Urine (24) hours
vitro quantitative, diagnostic determination of urea in human serum immediately with plenty of soap and water. In case of severe injuries; Normal & abnormal commercial control serum of known Urea : 20-35 g/24hrs (330-580 mmol/24hrs)
or urine on both automated and manual applications. seek medical advice immediately. concentrations should be analyzed with each run. BUN : 9.3-16.4 g/24hrs
Background Reagent (R) contains sodium azide which may react with copper or Performance Characteristics Spectrum Diagnostics does not interpret the results of a
Urea is the major product of protein nitrogen metabolism. It is lead plumbing. Precision clinical laboratory procedure;interpretation of the results is
synthesized by the urea cycle in the liver and excreted through Within run (Repeatiblity) considered the responsibility of qualified medical personnel.
the kidneys. The circulating levels of urea depend upon protein Reagent Storage and Stability All indications of clinical significance are supported by
Spectrum Urea UV Single reagent is supplied ready-to-use and Level 1 Level 2
intake, protein catabolism and kidney function. Elevated urea levels literature references.
can occur due to renal impairment or in some diseases such as stable up to the expiry date labeled on the bottles, Once opened, n 20 20
diabetes, infection, congestive heart failure and during different liver the opened vial is stable for 3 months at the specified temperature.
Mean (mg/dL) 45 150 Analytical Range
diseases. Determination of blood urea nitrogen is the most widely
used screening test for renal function together with serum creatinine. Deterioration SD 0.7 2.7 0.9 – 300 mg/dL (0.15 - 49.8 mmol/L).
Do not use Spectrum Ultimate Urea reagent if it is turbid or if the CV% 1.5 1.95
Method absorbance of the working reagent is less than 0.9 at 340 nm. Waste Disposal
urease-UV fixed rate (enzymatic method). Failure to recover control values within the assigned range may be This product is made to be used in professional laboratories. Please
an indication of reagent deterioration. consult local regulations for a correct waste disposal.
Run to run (Reproducibility) S56: dispose of this material and its container at hazardous or
Assay Principle
Specimen Collection and Preservation Level 1 Level 2 special waste collection point.
The series of reactions involved in the assay are as follows :
No special preparation of the patient is required. Use nonhemolyzed S57: use appropriate container to avoid environmental
n 20 20
serum or plasma only.The only acceptable anticoagulants are contamination.
1. Urea is hydrolyzed in the presence of water and urease to Mean (mg/dL) 47 153
heprin, EDTA and fluoride. Do not use ammonium heparin plasma. S61: avoid release in environment. refer to special instructions/
produce ammonia and carbon dioxide.
Stability:
o
7 days at 15 –25 C ; 7 days at 2 – 8 C;
o
SD 0.82 2.81 safety data sheets.
o
1 year at -20 C
Urease CV% 1.63 2.15 Referances
Urea + H2O 2NH3 + CO2 Urine samples are prediluted 1 : 50 with ammonium free water prior
to assay. 1. Batton, C. J & Crouch, S.R: Anal. Chem. , 1977, 49:464-469 .
Stability:
o
2 days at 15 – 25 C ; 7 days at 2 – 8 C;
o Methods Comparison 2. Shephard MD, Mezzachi RD: Clin Biochem Revs, 4:61-7, 1983.
2. In the presence of glutamate dehydrogenase (GLDH) and
1 month at -20 C
o A comparison between Spectrum Diagnostics Urea (UV) reagent 3. Tiffany TO, jansen JM, Burtis CA,Overtion JB, SCOTT CD.
reduced nicotinamide adenine dinucleotide (NADH),the
and a commercial reagent of the same methodology was performed Enzymatic kinetic rate and end point analyses of substrate, by
ammonia combines with a-ketoglutarate (a-KG) to produce
on 20 human sera. A correlation of 0.992 was obtained. use of a gemsaec fast analyzer. Clin Chem. 1972;18:829-840.
L-glutamate. System Parameters
Wavelength 340 nm 4. Tietz NW, Ed.Clinical guide to laboratory tests. 2ND.
Optical path 1 cm Sensitivity Philadelphia: WB Saunders;1990:566.
2NH4 + 2a-KG 2 L-Glutamate When run as recommended, the minimum detection limit of the
GLDH + Assay type Fixed Rate
+ assay is 0.9 mg/dL.
+
2 NAD + H2O Direction Decrease
2 NADH
e.g.: Reagent volume 1 ml Linearity ORDERING INFORMATION
The rate decrease in the NADH concentration is directly proportional Sample volume 10 ml The reaction is linear up to a urea concentration of 300 mg/dL. CATALOG NO QUANTITY
to the urea concentration in the specimen. It is determined by First read time 30 seconds Specimens showing higher concentration should be diluted 1+2
with physiological saline and repeat the assay (result × 3). 320 001 2 x 20 ml
measuring the absorbance at 340 nm. Delay time 60 seconds
320 002 6 x 20 ml
Last read time 90 seconds
o
Reagents Temperature 37 C
320 004 2 x 100 ml
Standard urea (ST) 107 mg/dL Zero adjustment Against Air Serum, plasma
Reagent High 2.0 AU Haemolysis
Tris Buffer ( pH 8.5) 50 mmol/L Sensitivity 0.9 mg/dL (0.15 mmol/L) Erythrocyte contamination doesn’t elevate results.Haemolytic
a-Ketoglutarate 10 mmol/L Linearity 300 mg/dL (49.8 mmol/L) specimens may cause high absorbance flagging.
GLDH 8.0 K U/L
Urease 5.0 K U/L Procedure Icterus
Sodium azide 8.0 mmol/L No significant interference.
Standard Specimen
NADH >0.20 mmol/L
Sodium azide 8 mmol/L Reagent (R) 1 ml 1 ml Lipemia
Standard 10 ml ------ Lipemic specimens may cause high absorbance flagging
The reagent also contains additives required to maintain NADH in Specimen ------ 10 ml Diluted sample treatment may be recommended.
its reduced forrm.
Mix, and after 30 seconds read the absorbance A1 of the standard Anticoagulants
For further information, refer to the Urea/Bun reagent material safety or specimen. Exactly 1 minute later, read the absorbance A2 of Ammonium heparin should not be used.
data sheet. standard or specimen.
Others
Reagent Preparation Ammonium ions should be avoided since it may cause
erroneously elevated results.
Spectrum Ultimate Urea reagent is supplied ready-to-use and stable
up to the expiry date labeled on the bottles. Once opened, the
o
opened vial is stable for 3 months at 2 - 8 C.
66 67
Uric acid - Liquizyme SYMBOLS IN PRODUCT LABELLING
Serum blank: Extremely lipemic samples may give falsely elevated
results and a serum blank must be run. Add 20 ml serum to 1 ml
Expectd Values
Child 2.0 -5.5 mg/dL (0.119 – 0.327 mmol/L)
Uricase-PAP (Single Reagent) EC REP Authorised Representative Temperature Limitation water. Zero the spectrophotometer with water.Read and record
IVD For in-vitro diagnostic use Use by/Expiration Date absorbance and subtract reading from test absorbance. Adult male 3.5 -7.2 mg/dL (0.208 – 0.428 mmol/L)
REF: 323 001 (4 x 30 ml) 120 test LOT Batch Code/Lot number CAUTION. Consult instructions Adult female 2.6 -6.0 mg/dL (0.155-0.357 mmol/L)
REF: 323 002 (4 x 50 ml) 200 test Catalogue Number for use Quality Control
REF
REF: 323 003 (4 x 100 ml) 400 test Normal & abnormal commercial control serum of known Urine 250 -750 mg/day (14.8-44.6 mmol/day)
REF: 323 004 (4 x 60 ml) 240 test concentrations should be analyzed with each run.
REF: 323 005 (2 x 500 ml) 1000 test Spectrum Diagnostics does not interpret the results of a
REF: 323 006 (4 x 250 ml) 1000 test Precautions and Warnings Performance Characteristics clinical laboratory procedure; interpretation of the results is
Do not ingest or inhalate. In case of contact with eyes or skin; rinse considered the responsibility of qualified medical personnel.
Precision
Intended Use immediately with plenty of soap and water. In case of severe injuries; All indications of clinical significance are supported by
Spectrum Diagnostics liquizyme uric acid reagent is intended for the seek medical advice immediately. literature references.
in-vitro quantitative, diagnostic determination of uric acid in human
serum or urine on both manual and automated systems. Reagent Storage and Stability Level 1 Level 2
All reagents are stable until expiration date stated on label when n 20 20 Analytical Range
o
Background stored refrigerated at 2 - 8 C. 1.0 – 20 mg / dL ( 0.06 – 1.19 mmol/l).
Uric acid is the end product of purine metabolism. Nearly half of Mean (mg/dL) 4.46 11.42
the uric acid is eliminated and replaced daily by way of urinary SD 0.15 0.21 Waste Disposal
excretion and through microbial degradation in the intestinal tract. Deterioration This product is made to be used in professional laboratories.
CV% 3.38 1.88
Hyperuricaemia may be observed in renal dysfunction, gout, Uric Acid Single reagent is normally clear or pale pink. Do not use Please consult local regulations for a correct waste disposal.
leukemia, polycythaemia, atherosclerosis, diabetes,hypothyroidism, liquizyme uric acid reagent if it is turbid or if the absorbance is S56: dispose of this material and its container at hazardous or
or in some genetic diseases. Decreased levels are present in greater than 0.15 AU at 546 nm. special waste collection point.
patients with Wilson’s disease, bronchogenic carcinoma, severe S57: use appropriate container to avoid environmental
hepatocellular disease and Hodgkin’s disease. Specimen Collection and Preservation Level 1 Level 2 contamination.
The only acceptable anticoagulants are heparin and EDTA. Uric n 20 20 S61: avoid release in environment. refer to special instructions/
Method acid in serum & plasma samples remains stable for 3 days at room safety data sheets.
Uricase-POD enzymatic colorimetric method with 4-amino-
o
temperature; 3 to 5 days if kept at 4oC and for 6 months at -20 C. Mean (mg/dL) 4.51 11.59
antipyrine. Urine samples should be diluted 1:10 before assay with physiological SD 0.23 1.32 References
saline. It is recommended that 15 ml of sodium hydroxide 2 mol/l, be
added to the urine samples to keep urine alkaline and prevent ureate CV% 3.46 1.97
Assay Principle 1. Barham D.and Trinder P., Analyst 97,142-145 (1972).
The assay is based upon the methods of modified trinder peroxidase percipitation. upon receipt, urine sample pH should be checked and 2. Fossati P.,Prencipe L.,and Berti G., Clin. Chem . 26/2,227-273
assay using 3,5-dichloro-2-hydroxybenzenesulfonic acid (DCHB) kept over 8.0. Methods Comparison (1980).
(2). The series of the reactions involved in the assay system is as A comparison between Spectrum Diagnostics Uric Acid reagent and 3. Richterich R, colombo JP. Klinische Chemie. 4th ed.basel:karger
follows: System Parameters a commercial reagent of the same methodology was performed on s;1978 :319-324 .
20 human sera. A correlation of 0.979 was obtained. 4. Tiffany to, jansen JM, Burtis CA,Overton JB, scott
1. Uric acid is oxidized to allantoin by uricase with production of Wavelength Hg 546 nm (500 – 550 nm) cd.Enzymatic kinetic rate and end point analyses of substrate,
hydrogen peroxide. Optical path 1 cm Sensitivity by use of a GEMSAEC fast analyzer. Clin Chem. 1972; 18 :
Assay type Endpoint When run as recommended, the minimum detection limit of this 829-840.
Uric acid Allantoin Direction Increase assay is 1 mg/dL (0.06 mmol/L). 5. Tietz NW, ED. Clinical guide to laboratory tests. 2nd ED.
+ Uricase + Sample : Reagent Ratio 1 : 50 philadelphia: WB Sauners; 1990: 566.
O2 + H2O CO2 + H2O2 e.g. : Reagent volume 1 ml Linearity
Sample volume 20 ml The reaction is linear up to a uric acid concentration of 20 mg/dl.
o o
2. the peroxide react with 4-amino-antipyrine and (DCHB) in Temperature 37 C or 15 – 25 C Specimens showing higher concentration should be diluted 1+1
o ORDERING INFORMATION
the presence of peroxidase to yield a quinoneimine dye. Reaction Time 5 minutes at 37 C using physiological saline, reassayed and the result multiplied by
o
The subchange in absorbance at 546 nm (500-550 nm) is 10 minutes 15 – 25 C two. CATALOG NO QUANTITY
proportional to uric acid concentration in the sample. Zero adjustment Reagent blank
323 001 4 x 30 ml
Reagent Blank Limits Low 0.00 AU Interfering Substance 323 002 4 x 50 ml
Quinoneimine High 0.15 AU Haemoglobin
H2O2 +Phenol POD 323 003 4 x 100 ml
+ + Sensitivity 1.0 mg/dL (0.06 mmol/L) No interference up to a haemoglobin level of 200 mg/dl. 323 004 4 x 60 ml
H 2O Linearity 20 mg/dl (1.19 mmol/L)
4-AAP + DCHB 323 005 2 x 500 ml
Icterus 323 006 4 x 250 ml
Procedure No significant interference from free bilirubin up to a level of 8mg/dl
Reagents
Blank Standard Specimen and from conjugated bilirubin up to a level of 12mg/dl.
Standard uric acid (ST)
6 mg/dL 0.357 mmol/L Reagent (R) 1.0 ml 1.0 ml 1.0 ml Lipemia
Standard ------ 20 µl ------ No significant interference with mild to moderate lipemia.
Reagent (R)
Phosphate Buffer 100 mmol/L Specimen ------ ------ 20 µl Drugs
(DCHB) 5.0 mmol/lL
Of the drugs tested in vitro,methyldopa and noramidopyrine cause
Potassium hexacyanoferrate 80 mmol/L o
Mix and incubate for 5 minutes at 37 C or 10 minutes at artificially Low uric acid values at the tested drug Level.
4-amino-antipyrine 0.6 mmol/lL
15 –25 C .Measure absorbance of specimen (Aspecimen) and
o
Peroxidase >3000 U/L

standard (Astandard) against reagent blank within 30 minutes. Others
Uricase > 500 U/L
Physiological ascorbic acid concentration does not interfere with
the test. Ascorbic Acid levels higher than 170 mmol/l (3.0 mg/dl)
For further information, refer to the Uric acid reagent material safety Calculation
decreases the apparent uric acid concentration significantly.
data sheet. (Aspecimen)
Serum uric acid concentration (mg/dL) = x6
(Astandard)
Reagent Preparation
Spectrum Uric acid liquizyme Single reagent is supplied ready-to- (Aspecimen)
use and stable up to the expiry date labeled on the bottles when Concentration of uric acid in urine = x 6 x 10
o (Astandard)
properly stored refrigerated at 2 – 8 C. Once opened, the opened
vial is stable for 3 months at the specified temperature.
68 69
ii-Enzymes
70 71
Acid Phosphatase (ACP) SYMBOLS IN PRODUCT LABELLING
LINEARITY:
The reaction is linear up to ACP concentration 74 U/L,
(Colorimetric Test EC REP Authorised Representative Temperature Limitation Specimen Shows higher concentration should be diluted 1+3 with
For in-vitro diagnostic use Use by/Expiration Date
with α-Naphthylphosphate) IVD
physiological saline and repeat assay (Result x 3).

Expected Values
REF 229 001 50 Tests Total Acid Phosphatase
Men £ 4,7 U/l
R1 Substrate 5 x10 ml Women £ 3,7 U/l
R2 For Total ACP 1 x 50 ml Prostatic Acid Phosphatase £ 1,6 U/l
R3 For Non-Prostatic ACP 1 x 50 ml
R4 Stabilizer 1 x 2 ml LITERATURE
1.Hillmann G. Z.Klin.Chem.Klin.Biochem.9, p.273.
METHOD / REACTION PRINCIPLE: Stability of Working reagents

o
α-Naphthylphosphate is hydrolysed by ACP to phosphate and 3 days at 2 - 8 C
o
α-naphthol, which is converted with FRTR-salt into an azo dye. The 1 day at 18 - 25 C
increase of absorbance at 405 nm is proportional to the total ACP
activity in the sample. The prostatic acid phosphatase (PACP) can SAMPLES :
be blocked by tartrate and can be determined indirectly (through the Serum, no plasma! Avoid hemolysis!
non-prostatic ACP) by calculation of the activity difference. Use immediately or stabilize :
Add 1 drop of 0.1% acetic acid to 1 ml of serum:
ACP o
α-Naphthylphosphate + H2O phosphate + 1-naphthol ACP is stable for 3 days at 2-8 C .
α-Naphthol + FRTR-salt Azo dye PROCEDURE:
REAGENTS: (Concentrations in the test) I-Manual assay for Total ACP
Reagent A 1.0 ml (Pre-heated At 37°C)

R1 : 1-Naphthyl phosphate 10 mmol/l Sample 100 µl
Fast Red TR-salt 1.5 mmol/l
(4-chloro-2-methylphenyl diazonium salt) Mix, Wait 5 minutes then measure absorbance (at 405 nm) each one
R2 : Citrate buffer pH 5.2 100 mmol/l minute for 3 minutes. Determine ΔA /min
R3 : Citrate buffer pH 5.2 100 mmol/l
Tartrate 135 mmol/l Calculation: ΔA /min x 743 = TACP Activity in sample
R4 : Stabilizer ( Acetic acid) 0.8 mol/l
The sealed reagents are stable up to the indicated expiry date

o II-Manual assay for Non-Prostatic ACP
if stored at 2 - 8 C.
Reagent B 1.0 ml (Pre-heated At 37°C)
PREPARATION AND STABILITY OF Sample 100 µl
WORKING REAGENTS
Mix, Wait 5 minutes then measure absorbance (at 405 nm) each one
R2 and R3 are ready for use minute for 3 minutes. Determine ΔA /min
Reagent A (determination of Total ACP):
Dissolve the contents of R1 (substrate) in Calculation: ΔA /min x 743 = NPACP Activity
10 ml of buffer solution R2.
Mark label with „A“. CALIBRATORS AND CONTROLS
For the calibration of automated analyzers Spectrum Multicalibrator
Reagent B (determination of Non-Prostatic is recommended, for quality control use Spectrum Normotrol and
ACP): Dissolve the contents of R1 (substrate) pathotrol.
in 10 ml of tartrate solution R3.Mark label withB
72 73
Alkaline phosphatase (ALP) SYMBOLS IN PRODUCT LABELLING
Precision
Waste Disposal
Liquizyme (9 + 1) IFCC EC REP Authorised Representative
Use by/Expiration Date Within run (Repeatiblity)
E.C.3.1.3.1. LOT
REF
Batch Code/Lot number
Catalogue Number
for use
Level 1 Level 2

S57:
special waste collection point.
use appropriate container to avoid environmental
Consult instructions for use Manufactured by n 20 20 contamination.
REF: 214 001 ( 4 x 20 ml) 80 test
REF: 214 002 (10 x 10 ml) 100 test Mean (U/L) 177.7 359.7
safety data sheets.
REF: 214 003 ( 9 x 20 ml) 180 test Reagent Storage and Stability SD 1.71 1.5
REF: 214 004 ( 4 x 60 ml) 240 test All reagents are stable until expiration date stated on label when
REF: 214 005 ( 5 x 20 ml) 100 test o
stored refrigerated at 2 - 8 C. CV% 0.96 0.43 References
o
Working solution is stable for 4 weeks at 2 – 8 C or 5 days at
Intended Use o 1. Moss DW. Alkaline phosphatase isoenzymes. Clin Chem.
15 - 25 C.
Spectrum Diagnostics liquizyme Alkaline Phosphatase reagent is Run to run (Reproducibility) 1982;28:2007-2016 .
intended for the in-vitro quantitative, diagnostic determination of Deterioration Level 1 Level 2 2. Moss DW, Henderson AR, Kachmar JF. Enzymes in:Tietz NW,
ALP in human serum on both automated and manual systems. Do not use liquizyme ALP reagent if it is turbid or if the absorbance ed. Fundamentals of clinical chemistry. 3 rd ed. Philadelphia:
n 20 20
of the working reagent is more than 1.0 at 405 nm. Failure to recover WB Saunders; 1987:346-421.
Background control values within the assigned range may be an indication of Mean (U/L) 178.5 365.5
Alkaline phosphatase (ALP) catalyzes the hydrolysis of a wide 3. Tietz NW, Rinker AD, Shaw LM. IFCC methods for the
reagent deterioration. SD 1.82 1.86
variety of physiologic and non-physiologic phosphoric acid esters measurement of catalytic concentration of enzymes. Part 5.
in alkaline medium (pH optimum 10). The liver and biliary tract are CV% 1.15 0.55 IFCC method for alkaline phosphatase . J Clin Chem Clin
Specimen Collection and Preservation Biochem. 1983;21:731-748.
the source of alkaline phosphatase in normal sera. Normal alkaline
Serum and Plasma
phosphatase levels are age dependent being higher in children
Nonhaemolyzed fresh serum is the preferred specimen. Heparin is Methods Comparison 4. Zawta B, Klein G, Bablok W. Temperaturumrechnung in der
and adolescents in comparison to adults. ALP is one of the tests of A comparison between Spectrum Diagnostics ALP reagent and a Klinischen Enzymologie? Klin lab. 1994:40:23-32. Sensitivity
the only acceptable anticoagulant. Complexing anticoagulants such
choice for evaluating cholestasis and obstructive juandice. Elevated commercial reagent of the same methodology was performed on 20
as citrate, oxalate, and EDTA must be avoided.
levels are found in many diseases including hepatitis, cirrhosis, human sera. A correlation of 0.988 was obtained.
Alkaline phosphatase activity may slowly increase in serum samples
malignancy, and in bone diseases.
stored at room temperature. Previously frozen or lypholized sera
Sensitivity ORDERING INFORMATION
may show a marked decrease in values immediately upon thawing
Method or reconstitution.The activity then increases to the initial values, and When run as recommended, the minimum detection limit of this CATALOG NO QUANTITY
Kinetic method according to the International Federation of Clinical the rate of this increase is time and temperature dependent. assay is 5.0 U/L.
Chemistry (IFCC) (3). 214 001 4 x 20 ml
214 002 10 x 10 ml
Stability:
o
2 months at -20 C ; 4 weeks at 4 – 8 C;
o
Linearity
214 003 9 x 20 ml
Assay Principle o
7 days at 20 – 25 C The reaction is linear up to alkaline phosphatase concentration of
214 004 4 x 60 ml
Alkaline phosphatase (ALP) hydrolyzes p-Nitrophenylphosphate 750 U/L; specimens showing higher concentration should be diluted
214 005 5 x 20 ml
(p-NPP) to p-Nitrophenol and phosphate. System Parameters 1+5 with physiological saline and repeat the assay (result × 6).
Wavelength 405 nm (400 – 420 nm)
p-Nitrophenylphosphate + H2O ALP p-Nitrophenol+ Phosphate Optical path 1 cm Interfering Substances
Assay type Kinetic Serum, plasma
The increase of absorbance per minute at 405 nm is proportional to Direction Increase
the enzyme activity. Sample : Reagent Ratio 1 : 100 Haemolysis
e.g.: Reagent volume 1 ml A 200 mg/dL haemoglobin results in a 10 % negative bias.
Reagents Sample volume 10 ml
o o
Temperature 37 C or 30 C Icterus
Reagent 1 (R1 Buffer) Equilibration time 1 minute No significant interference up to bilirubin level of 40 mg/dL.
2-Amino-2-Methyl-1-Propanol (pH 10.3) 2.0 mol/L Read time 1 to 3 minutes
Zero adjustment Against air Lipemia
MgCl2 2.0 mmol/L No significant interference from lipemia up to 1000 mg/dL.
Reagent 2 (R2 Substrate) High 1.0 AU Expected Values
p-Nitrophenylphosphate 16 mmol/L Sensitivity 5 U/L o
30 C 37 C
o
Linearity 750 U/L

Males (20 - 50) years 30 - 90 U/L 53-128 U/L
For further information, refer to the Alkaline phosphatase reagent
material safety data sheet. Procedure Males (≥ 60) years 30 - 90 U/L 56-119 U/L
Pipette in a test tube: Females (20 - 50) years 20 - 80 U/L 42-98 U/L
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Working 1.0 ml Females (≥ 60) years 40 - 111 U/L 53-141 U/L
immediately with plenty of soap and water. In case of severe injuries; solution
Childern (1 - 12) years ≤ 350 U/L ≤ 460 U/L
seek medical advice immediately. Specimen 10 ml
o
Temperature conversion factor is 1.22 (25 30 C ) and
Reagent Preparation Mix, read initial absorbance after 1 minute. and start timer 1.52 (25
o
37 C ).
Prepare working solution as following: simultaneously. Read again after 1, 2 and 3 minutes.
Determine the mean absorbance change per minute (DA/min).
REF: 214 001: add 2 ml from R2 to one bottle of R1; mix gently.
REF: 214 002: add 1 ml from R2 to one bottle of R1; mix gently. Calculation considered the responsibility of qualified medical personnel.
REF: 214 003: add 2 ml from R2 to one bottle of R1; mix gently. To calculate the alkaline phosphatase (ALP) activity, Use the All indications of clinical significance are supported by
REF: 214 004: add 6 ml from R2 to one bottle of R1; mix gently. following formula U/L = 5454 ×DA 405 nm/min literature references.
REF: 214 005: add 2 ml from R2 to one bottle of R1; mix gently.
Quality Control
Or prepare the working solution according to the number of tests Normal & abnormal commercial control serum of known Analytical Range
required by mixing 9 volumes of reagent 1 (R1) and 1volume of concentrations should be analyzed with each run. 5 – 750 U/L.
reagent 2 (R2),e.g. 900 ml R1 + 100 ml R2.
74 75
Alkaline phosphatase (ALP) SYMBOLS IN PRODUCT LABELLING
Precision
Liquizyme (1 + 1) IFCC EC REP Authorised Representative
E.C.3.1.3.1. LOT
REF
Catalogue Number
for use
Level 1 Level 2 literature references.
Consult instructions for use Manufactured by n 20 20

Mean (U/L) 177.7 359.7 Analytical Range
REF: 215 001 (2 x 25 ml) 50 test 5 – 750 U/L.
SD 1.71 1.5
REF: 215 002 (4 x 25 ml) 100 test
CV% 0.96 0.43 Waste Disposal
Intended Use Specimen Collection and Preservation This product is made to be used in professional laboratories.
Spectrum Diagnostics liquizyme Alkaline Phosphatase reagent is Please consult local regulations for a correct waste disposal.
Serum and Plasma S56: dispose of this material and its container at hazardous or
intended for the in-vitro quantitative, diagnostic determination of ALP Run to run (Reproducibility)
Nonhaemolyzed fresh serum is the preferred specimen. Heparin
in human serum on both automated and manual systems. special waste collection point.
is the only acceptable anticoagulant. Complexing anticoagulants Level 1 Level 2
such as citrate, oxalate, and EDTA must be avoided. Alkaline n 20 20
Background contamination.
phosphatase activity may slowly increase in serum samples stored
Alkaline phosphatase (ALP) catalyzes the hydrolysis of a wide Mean (U/L) 178.5 365.5 S61: avoid release in environment. refer to special instructions/
at room temperature. Previously frozen or lyophilized sera may
variety of physiologic and non-physiologic phosphoric acid esters safety data sheets.
show a marked decrease in values immediately upon thawing or SD 1.82 1.86
in alkaline medium (pH optimum 10). The liver and biliary tract are recon-stitution.The activity then increases to the initial values, and
the source of alkaline phosphatase in normal sera. Normal alkaline the rate of this increase is time and temperature dependent. CV% 1.15 0.55 References
phosphatase levels are age dependent being higher in children
and adolescents in comparison to adults. ALP is one of the tests of Stability:
o
2 months at – 20 C ; 4 weeks at 4 – 8 C;
o
Methods Comparison 1. Moss DW. Alkaline phosphatase isoenzymes. Clin
choice for evaluating cholestasis and obstructive juandice. Elevated o
7 days at 20 – 25 C A comparison between Spectrum Diagnostics ALP reagent and a Chem.1982;28:2007-2016.
levels are found in many diseases including hepatitis, cirrhosis, commercial reagent of the same methodology was performed on 20
malignancy, and in bone diseases. System Parameters human sera. A correlation of 0.990 was obtained. 2. Moss DW, Henderson AR, Kachmar JF. Enzymes in: Tietz
Wavelength 405 nm (400 – 420 nm) NW, ed. Fundamentals of Clinical Chemistry. 3 rd ed.
Method Optical path 1 cm Sensitivity Philadelphia: WB Saunders; 1987:346-421.
Kinetic method according to the International Federation of Clinical Assay type Kinetic When run as recommended, the minimum detection limit of this 3. Tietz NW, Rinker AD, Shaw LM. IFCC methods for the
Chemistry (IFCC) (3). Direction Increase assay is 5.0 U/L. measurement of catalytic concentration of enzymes. Part5.
Sample : Reagent Ratio 1 : 100 IFCC method for alkaline phosphatase . J Clin Chem Clin
Assay Principle e.g.: Reagent volume 1 ml Linearity Biochem.1983;21:731-748.
Alkaline phosphatase (ALP) hydrolyzes p-Nitrophenylphosphate Sample volume 10 ml The reaction is linear up to alkaline Phosphatase concentration of
(p-NPP) to p-Nitrophenol and phosphate. Temperature 37 C or 30 C
o o
750 U/L; specimens showing higher concentration should be diluted 4. Zawta B, Klein G, Bablok W. Temperaturumrechnung in der
Equilibration time 1 Minute 1+5 with physiological saline and repeat the assay (result×6). Klinischen Enzymologie? Klin lab. 1994:40:23-32.
ALP
p-Nitrophenylphosphate + H2O p-Nitrophenol + Phosphate Read time 1 to 3 minutes
Zero adjustment Against air Interfering Substances
The increase of absorbance per minute at 405 nm is proportional to Reagent Blank Limits Low 0.2 AU Serum, plasma ORDERING INFORMATION
the enzyme activity. High 1.0 AU
Sensitivity 5 U/L Haemolysis CATALOG NO QUANTITY
Reagents Linearity 750 U/L A 200 mg/dL haemoglobin results in a 10 % negative bias. 215 001 2 x 25 ml
215 002 4 x 25 ml
Reagent 1 (R1 Buffer) Procedure Icterus
2-Amino-2-Methyl-1-Propanol (pH 10.3) 2.0 mol/L Pipette in a test tube: No significant interference up to bilirubin level of 40 mg/dL.
MgCl2 2.0 mmol/L Working 1.0 ml ( or add 0.5 ml R1 + 0.5 ml R2 ) Lipemia

solution No significant interference from lipemia up to 1000 mg/dL.
Reagent 2 (R2 Substrate)
Specimen 10 ml
p-Nitrophenylphosphate 16 mmol/L Expected Values
o o
Mix, read initial absorbance after 1 minute. and start timer 30 C 37 C
For further information, refer to the Alkaline phosphatase reagent simultaneously. Read again after 1, 2 and 3 minutes. Determine the
material safety data sheet. Males (20 - 50) years 30 - 90 U/L 53 - 128 U/L
mean absorbance change per minute (DA/min).
Males (> 60) years 30 - 90 U/L 56 - 119 U/L
Precautions and Warnings Calculation Females (20 - 50) years 20 - 80 U/L 42 - 98 U/L
Do not ingest or inhalate. In case of contact with eyes or skin; rinse To calculate the alkaline phosphatase (ALP) activity. Use the
immediately with plenty of soap and water. In case of severe injuries; following formulae U/l = 5454 × DA 405 nm /min Females (> 60) years 40 - 111 U/L 53 - 141 U/L
seek medical advice immediately. Childern (1 - 12) years < 350 U/L <460 U/L
Quality Control
Reagent Preparation Storage and Stability Normal & abnormal commercial control serum of known Temperature conversion factor is 1.22 (25
o
30 C ) and
All reagents are stable until expiration date stated on label when concentrations should be analyzed with each run. 1.52 (25
o
37 C ).
o
stored refrigerated at 2 - 8 C .
Working solution can be prepared by adding equal volumes
o o
from R1 and R2; Stability: 1 month at 2 – 8 C or 5 days at 15-25 C.
Deterioration
Do not use liquizyme ALP reagent if it is turbid or if the absorbance
of the working reagent is more than 1.0 at 405 nm. Failure to recover
control values within the assigned range may be an indication of
reagent deterioration.
76 77
ALKALINE PHOSPHATASE SYMBOLS IN PRODUCT LABELLING
Expected Values
Single Reagent EC REP Authorised Representative
37 C
o
(Kinetic - IFCC method) LOT Batch Code/Lot number CAUTION. Consult instructions Males (20 - 50) years 53-128 U/L
REF Catalogue Number for use Males (> 60) years 56-119 U/L
Females (20 - 50) years 42-98 U/L
REF: 217 002 ( 4 x 25 ml) 100 test
REF: 217 003 ( 4 x 50 ml) 200 test Females (> 60) years 53-141 U/L
Childern (1 - 12) years <460 U/L
Intended Use Stability:
o
2 months at -20 C ; 4 weeks at 4 – 8 C;
o
o
Spectrum Diagnostics liquizyme Alkaline Phosphatase reagent is 7 days at 20 – 25 C The reference values are to be considered as indicative only.
intended for the in-vitro quantitative, diagnostic determination of Every Laboratory should establish its own normal ranges.
ALP in human serum on both automated and manual systems. System Parameters
Background Optical path 1 cm Spectrum Diagnostics does not interpret the results of a
Alkaline phosphatase (ALP) catalyzes the hydrolysis of a wide Assay type Kinetic clinical laboratory procedure;interpretation of the results is
variety of physiologic and non-physiologic phosphoric acid esters Direction Increase considered the responsibility of qualified medical personnel.
in alkaline medium (pH optimum 10). The liver and biliary tract are Sample : Reagent Ratio 1 : 100 All indications of clinical significance are supported by
the source of alkaline phosphatase in normal sera. Normal alkaline e.g.: Reagent volume 1 ml literature references.
phosphatase levels are age dependent being higher in children Sample volume 10 ml
o
and adolescents in comparison to adults. ALP is one of the tests of Temperature 37 C
choice for evaluating cholestasis and obstructive juandice. Elevated Interval time 30 Sec.
levels are found in many diseases including hepatitis, cirrhosis, Delay/Lag time 60 Sec.
malignancy, and in bone diseases. Measurement Against distalled Water Analytical Range
5 – 750 U/L.
Method Reagent Blank Limits Low 0.2 AU
Kinetic method according to the International Federation of Clinical High 1.0 AU Waste Disposal
Chemistry (IFCC) Sensitivity 5 U/L This product is made to be used in professional laboratories.
o
Linearity at 37 C 750 U/L Please consult local regulations for a correct waste disposal.
Assay Principle S56: dispose of this material and its container at hazardous or
p-Nitrophenyl phosphate is converted to p-Nitrophenol and special waste collection point.
phosphate by alkaline phosphatase. The increase of absorption at Procedure S57: use appropriate container to avoid environmental
405 nm is proportional to the alkaline phosphatase concentration in contamination.
Pipette in a test tube:
the sample. S61: avoid release in environment. refer to special instructions/
Reagent (R) 1.0 ml safety data sheets.
Reagents
Specimen 10 ml References
Reagent (R)
o
Mix well and Incubate at 37 C for 60 sec. 1. Moss DW. Alkaline phosphatase isoenzymes. Clin Chem.
Substrate Reagent
Measure absorbance increase every 30 sec for 2 minutes and 1982;28:2007-2016 .
For further information, refer to the Aspartate aminotransferase determine the (DA/min). 2. Moss DW, Henderson AR, Kachmar JF. Enzymes in:Tietz NW,
Monoreagent material safety data sheet. ed. Fundamentals of clinical chemistry. 3 rd ed. Philadelphia:
Calculation WB Saunders; 1987:346-421.
Precautions and Warnings ALP Concentration (U/L) = DA/min x 5454
Do not ingest or inhalate. In case of contact with eyes or skin; rinse 3. Tietz NW, Rinker AD, Shaw LM. IFCC methods for the
immediately with plenty of soap and water. In case of severe injuries; Quality Control measurement of catalytic concentration of enzymes. Part 5.
seek medical advice immediately. Normal & abnormal commercial control serum of known IFCC method for alkaline phosphatase . J Clin Chem Clin
concentrations should be analyzed with each run. Biochem. 1983;21:731-748.
Reagent Preparation 4. Zawta B, Klein G, Bablok W. Temperaturumrechnung in der
Spectrum ALP-Single reagent is supplied ready-to-use. Sensitivity Klinischen Enzymologie? Klin lab. 1994:40:23-32. Sensitivity
Reagent Storage and Stability assay is 5.0 U/L.
The reagent is stable until expiration date stated on label when ORDERING INFORMATION
o
stored refrigerated at 2 - 8 C. Linearity
The reaction is linear up to alkaline phosphatase concentration of CATALOG NO QUANTITY
Deterioration 750 U/L 217 001 2 x 25 ml
Do not use liquizyme ALP reagent if it is turbid or if the absorbance 217 002 4 x 25 ml
of the working reagent is more than 1.0 at 405 nm. Failure to recover Interfering Substances 217 003 4 x 50 ml
control values within the assigned range may be an indication of Serum, plasma
Haemolysis
Specimen Collection and Preservation A 200 mg/dL haemoglobin results in a 10 % negative bias.
Serum and Plasma Icterus

Nonhaemolyzed fresh serum is the preferred specimen. Heparin is No significant interference up to bilirubin level of 40 mg/dL.
the only acceptable anticoagulant. Complexing anticoagulants such
as citrate, oxalate, and EDTA must be avoided. Lipemia
Alkaline phosphatase activity may slowly increase in serum samples No significant interference from lipemia up to 1000 mg/dL.
stored at room temperature. Previously frozen or lypholized sera
may show a marked decrease in values immediately upon thawing
or reconstitution.The activity then increases to the initial values, and
the rate of this increase is time and temperature dependent.
78 79
Alkaline Phosphatase - SYMBOLS IN PRODUCT LABELLING Calculation
EC REP Authorised Representative Use by/Expiration Date
Colorimetric IVD
LOT
For in-vitro diagnostic use
! CAUTION. Consult instructions
for use
OD serum sample – OD serum blank
xn
OD Standard
REF: 216 001 50 Test
Consult instructions for use (T) Toxic
Temperature Limitation n = 20 (Kind and king U/100 ml)
n = 142 (IU/L)

Quality Control
Intended Use Reagent 4 (R4 Color reagent) Normal & abnormal commercial control serum of known
Spectrum Diagnostics Alkaline Phosphatase Colorimetric reagent is Potassium ferricyanide 150 mmol/L concentrations should be analyzed with each run.
intended for the in-vitro quantitative, diagnostic determination of ALP
in human serum on both automated and manual systems. Precautions and Warnings Sensitivity
Do not ingest or inhalate. In case of contact with eyes or skin; rinse When run as recommended, the minimum detection limit of the
Background immediately with plenty of soap and water. In case of severe injuries; assay is 1 U/100mL.
Alkaline phosphatase (ALP) catalyzes the hydrolysis of a wide seek medical advice immediately.
variety of physiologic and non-physiologic phosphoric acid esters Linearity
in alkaline medium (pH optimum 10). The liver and biliary tract are Reagent preparation, Storage, and Stability For activities < 40 kind and king U/100 ml (285 IU/L) reassay using
the source of alkaline phosphatase in normal sera. Normal alkaline The reagents are supplied ready-to-use and stable up to the expiry a smaller volume such as 20 or 10 µl. multiply the result by 2.5 or 5
o
phosphatase levels are age dependent being higher in children date labeled on the bottles when stored at 2 – 8 C. respectively.
and adolescents in comparison to adults. ALP is one of the tests of
choice for evaluating cholestasis and obstructive juandice. Elevated Specimen Collection and Preservation Expected values
levels are found in many diseases including hepatitis, cirrhosis, Serum and Plasma
malignancy, and in bone diseases. Nonhemolyzed fresh serum is the preferred specimen. Heparin is Children: 10 – 20 Kind and king U/100 ml
the only acceptable anticoagulant. Complexing anticoagulants such 71 – 142 IU/L
Method as citrate, oxalate, and EDTA must be avoided.
ALP – (Colorimetric method). Alkaline Phosphatase activity may slowly increase in serum samples Adults: 3 – 13 kind and king U/100 ml
stored at room temperature. 21 – 92 IU/L
Assay Principle Stability:
o o
2 months at – 20 C; 4 weeks at 4 – 8 C;
Colorimetric determination of alkaline Phosphatase activity
o
7 days at 20 – 25 C Note
according to the following reaction: One kind and king unit is the amount of enzyme which in the given
System Parameters o
conditions liberates 1 mg of phenol in 15 minutes at 37 C
Phenylphosphate ALP Phenol + Phosphate Wavelength 510 nm ( Hg 492)
pH 10
Optical path 1 cm Waste Disposal
Assay type Endpoint This product is made to be used in professional laboratories.
phenol liberated is measured in the presence of 4-aminoantipyrine Direction Increase Please consult local regulations for a correct waste disposal.
o o
and Potassium ferricyanide. The presence of sodium arsenate in the Temperature 37 C and 20 – 25 C S56: dispose of this material and its container at hazardous or
reagent stops the enzymatic reaction. Zero adjustment Reagent Blank special waste collection point.
Reagents Procedure contamination.
Set up the following tubes S61: avoid release in environment. refer to special instructions/
Reagent 1 (R1 Buffer) pH 10 Serum Sample Serum blank Standard Reagent blank safety data sheets.
Disodium phenylphosphate 5.0 mmol/L
Carbonate-bicarbonate buffer 50 mmol/L R1 2ml 2ml 2ml 2ml References
o
Incubate for 5 minutes at 37 C 1. Kind Pra and King EJ: J clin patho 7: 322, 1954.
Reagent 2 (R2 Standard)
R2 ------ ------ 50 ml ------ 2. Marsh WH, Finger hut B, Kirsch E: clin chem. 5:119, 1959.
Phenol Equal to 20 kind and king U
Serum 50 ml ------ ------ ------ 3. Belfield A, GOLDBERG D.M – Enzyme 1971, 12,561.
Reagent 3 (R3 Blocking reagent) o
Incubate for exactly 15 minutes at 37 C
4-aminoantipyrine 60 mmol/L
Sodium arsenate 240 mmol/L R3 0.5 ml 0.5 ml 0.5 ml 0.5 ml
Buffer pH 10 Mix well or preferably vortex. CATALOG NO QUANTITY
Toxic reagent
R4 0.5 ml 0.5 ml 0.5 ml 0.5 ml 216 001 50 Test
R 45 : may cause cancer.
R 23/25 : toxic by inhalation and if swallowed. Serum ------ 50 ml ------ ------
S 28 : after contact with skin, wash immediately with plenty of
Dist.Water ------ ------ ------ 50 ml
water.
S 45 : in case of accident or if you feel unwell, seek medical Mix. Let stands for 10 minutes in the dark. Measure The color
advice immediately (show the label when possible). intensity is stable for 45 minutes.
80 81
Alpha Amylase SYMBOLS IN PRODUCT LABELLING
Expected Values (37°C)
(Single Reagent) - GALG2-CNP EC REP Authorised Representative

25°C 30°C 37°C
LOT Batch Code/Lot number CAUTION. Consult instructions Serum / Up to 55 U/L Up to 73 U/L Up to 100 U/L
Catalogue Number for use Plasma
REF: 219 001 2 x 25 ml REF
REF: 219 002 4 x 25 ml Consult instructions for use Manufactured by Random Up to 273 U/L Up to 365 U/L Up to 450 U/L
Urine
Specimens 24hrs Urine Up to 205 Up to 295 Up to 410
Background Serum, Heparinized plasma and Urine. U /24h U /24h U/24h
Measurements of amylase are used primarily in the diagnosis and
treatment of the diseases of the pancreas Amylase is found primarily Notes:
in the pancreas and salivary glands. When released in the digestive -- Saliva and skin contain alpha amylase: never pipette by mouth Interferences
tract, the enzyme hydrolyzes starch. Amylase determinations are and avoid skin contact with the reagents (use gloves). The following substances doesn’t interfere up to the concentration
useful in the diagnosis of diseases of the pancreas and parotids. -- Avoid use of haemolysed samples. of:
Elevated serum levels are associated with acute pancreatitis and -- The activity of alpha amylase in serum or plasma is stable for 7 Bilirubin conjugated 20 mg/dL
other pancreatic disorders as well as mumps and bacterial parotitis. days at +2-8°C, one month at –20°C. Bilirubin free 20 mg/dL
Hemoglobin 500 mg/dL
Method Procedure1 (Kinetic method) NaF 500 mg/dL
Kinetic method - GALG2-CNP Wavelength l = 405 nm Ascorbic acid 500 mg/dL
Light path 1 cm Glucose 5,0 g/dL
Assay Principle: Temperature 37°C Maltose 5,0 g/dL
Alpha amylase catalyzes the hydrolysis of 2-chloro-4-nitrophenyl-1- Measurement Against reagent blank
galactopyranosyl-maltoside (GALG2-CNP) to glucose polymers and Reaction Kinetic (increase) - Precision
p-nitrophenyl oligosaccaride at short chain producing 2-chloro-4- Within-run reproducibility
nitrophenol (CNP). Allow reagents to reach working temperature before use.
The increased extintion can be measured by spectrophotometry
at 405nm and results proportional at the activity of alpha amylase Within Standard-
Mean value CV
present in the sample. BLANK SAMPLE series deviation
[U/L] [%]
Reagent 1000 1000 mL n = 20 [U/L]
Reagents (concentrations in the test): Sample 1 77,7 1,009 1,298
Distilled water 25 ------ mL
Reagent (R) Sample ------ 25 mL Sample 2 196 1,834 0,935
Goods Buffer pH 6,0 50 mmol/L Sample 3 72,1 1,700 0,934

Mix thoroughly and incubate for 1 minute at 37°C.
GALG2-CNP 2.65 mmol/L Record initial absorbance and at 1 minute intervals thereafter for 3
Sodium chloride 300 mmol/L minutes against reagent blank. Between-run reproducibility
Calculate the difference between consecutive absorbance’s, and the
Calcium chloride 5 mmol/L Standard-
average absorbance difference per minute (DE/min). Day to day Mean value CV
deviation
Potassium thiocyanate 140 mmol/L n = 20 [U/L] [%]
[U/L]
Calculation:
EDTA 0,2 mmol/L Sample 1 79,5 0,934 1,176
Alpha amylase (U/L) - ∆E/min x 3060
Sample 2 195 0,674 0,345
(R) Reagent Contains potassium thiocyanate Procedure2 (Fixed rate colorimetric method)
Sample 3 72,5 0,934 1,288
X
- R 22: Harmful if swallowed
Harmful - S 36: Wear suitable protective clothing Wavelength l = 405 nm
Light path 1 cm
Temperature 37°C LINEARITY: 1500 U/L
Reagent Blank Against distilled water SENSITIVITY: 2 U/L
Reagent Storage and Stability MEASURED RANGE: 2 - 1500 U/L
Reaction Fixed rate (increase)
Spectrum amylase reagents are supplied ready-to-use and stable

up to the expiry date labeled on the bottles when properly stored References
o Allow reagents to reach working temperature before use.
refrigerated at 2 – 8 C. Once opened, the opened vial is stable for 5
months at the specified temperature. 1. Henry, R.J., Chiamori, N., Clin. Chem., 6;434, (1961)
Sample
2. Winn-Deen et Al., Clin. Chem. 24-10 (1989)
Reagent Preparation Reagent 1000 µl
The reagent is stable liquid ready to use 3. Lorentz, K., Clin. Chem. Clin. Biochem. 17,499 (1979)
Sample 25 µl
Precaution and warning
-- The present method describes the manual use of this kit. For Mix well and incubate exactly for 1 minute at 37°C then Read the
use with automatic analyzer see the specific applications. Absorbance A1.
After exactly 4 minutes later read the Absorbance A2. ORDERING INFORMATION
-- Presence of particulate material indicates deterioration of the
reagent. CATALOG NO QUANTITY
-- Do not use the reagent If the absorbance is > 0.600 at 405 nm. Calculation: 219 001 2 x 25 ml
-- Quality control data sheet of the reagent are available upon ∆E = A2 – A1 219 002 4 x 25 ml
request. Refer to the batch number on the label. Alpha amylase (U/L) = ∆E x 765
82 83
Creatine Kinase (CK) SYMBOLS IN PRODUCT LABELLING
Performance Characterstics
Precision
Liquid Reagent EC REP Authorised Representative
REF: 238 001 ( 6 x 5 ml) 30 Test LOT Batch Code/Lot number CAUTION. Consult instructions
Level 1 Level 2
REF: 238 002 ( 6 x 20 ml) 120 Test REF Catalogue Number for use
REF: 238 004 ( 6 x10 ml ) 60 Test Consult instructions for use Manufactured by n 20 20
Mean (U/L) 86 616
CV% 2.8 1.0
Intended Use Reagent preparation, Storage, and Stability
Spectrum Diagnostics Creatine Kinase (CK) reagent is intended for REF: 238 001 add 1 ml from R2 to one bottle of R1; mix gently
the in-vitro quantitative, diagnostic determination of Creatine kinase REF: 238 002 add 4 ml from R2 to one bottle of R1; mix gently Run to run (Reproducibility)
in human serum on both automated and manual systems. REF: 238 004 add 2 ml from R2 to one bottle of R1; mix gently
Level 1 Level 2
Background Or prepare the working solution according to the number of test n 20 20
Creatine kinase (CK) is an enzyme which is contained in heart, brain required by mixing 4 volumes of R1 with 1 volume of R2. Stability: 4
and skeletal muscles. Thus, an increase of circulating level of CK Mean (U/L) 77 624
weeks at 2-8oC away from light sources.
may be associated to myocardial infarction, acute cerebrovascular CV% 2.5 0.8
disease, trauma or diseases of skeletal muscles. After a myocardial Specimen Collection and Preservation
infarct, CK level begins raising between 4th and 6th hour after first Serum free of haemolysis or heparin plasma. Stability 2 days at Methods Comparison
acute symptoms, reaching the peak between 18th and 30th hour 20-25 oC, 7 days at 2-8oC, 4 weeks at -20oC protected from light. A comparison between Spectrum Diagnostics CK reagent and a
and coming back to normal values during the 3rd day. CK is present commercial reagent of the same methodology was performed on 20
in three different isoenzymatic forms, which could be separated by human sera. A correlation of 0.983 was obtained.
System Parameters
electrophoresis or column chromatography; each form is originated
Wavelength 340 nm (334-365 nm)
in different body tissues, paying off their diagnostic determinations. Sensitivity
Optical path 1 cm
The formula of present reagent is based on DGKC and IFCC When run as recommended, the minimum detection limit of the
Assay type Kinetic
recommendations. assay is 1 U/L.
Direction Increase
Sample: Reagent Ratio 1:25
Method e.g.: Reagent volume 1 ml Linearity
According to the recommendations of the International Federation of Sample volume 40 ml The reaction is linear up to CK concentration of 2000 U/l; specimens
Clinical Chemistry (IFCC). Temperature 37 C
o
showing higher concentration should be diluted 1+2 using
Equilibration Time 60 seconds physiological saline and repeat the assay (result×3).
Assay Principle Read time 1 to 3 minutes
Creatine kinase (CK) catalyzes the phosphorylation of ADP, in Zero adjustment against air Interferences:
the presence of creatine phosphate, to form ATP and creatine. Reagent blank limits No interferences were observed with haemoglobin until 5 g/L,
The catalytic concentration is determined from the rate of NADPH Sensitivity 1 U/L bilirubin 20 mg/dL and triglycerides 7 mmol/L. Other drugs and
formation, measured at 340 nm, by means of the hexokinase (HK) Linearity 2000 U/L substances may interfere3,4.
and glucose-6-phosphate dehydrogenase (G6PDH) coupled
Reactions1,2. Procedure References
CK 1. IFCC methods for the measurement of catalytic concentration
Creatine phosphate + ADP Creatine + ATP 1. Pipette into a thermostatized cuvette: of enzymes. Part 7: IFCC method for creatine kinase. JIFCC
HK Working solution 1 ml 1989; 1: 130-139.
ATP + Glucose ADP + Glucose-6-phosphate 2. Tietz Textbook of Clinical Chemistry, 3rd edition. Burtis CA,
+ G6PDH +
Serum 40 mL Ashwood ER. WB Saunders Co., 1999.
Glucose-6- phosphate+NADP 6-Phosphogluconate+NADPH +H
3. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed
2. Mix and incubate 60 seconds. AACC Press, 1995.
Reagents 4. Young DS. Effects of disease on Clinical Lab. Tests, 4th ed
Reagent 1 (pH 6.7) (Buffer / Coenzyme) 3. Read initial absorbance (A) of the sample, start the stopwatch
AACC 2001.
and read absorbance at 1 minute intervals thereafter for 3
Imidazol 125 mmol/L minutes.
D-Glucose 25 mmol/L ORDERING INFORMATION
N-Acetyl-L-Cysteine 25 mmol/L 4. Calculate the difference between absorbances and the
Magnesium acetate 12.5 mmol/L average absorbance differences per minute (DA/min). CATALOG NO QUANTITY
NADP 2.5 mmol/L 238 001 5 x 6 ml
EDTA 2 mmol/L 238 002 6 x 20 ml
Calculation
DA/min x 4127 = U/L CK 238 004 6 x 10 ml
Reagent 2 (Enzymes)
Units: One international unit (IU) is the amount of enzyme that
ADP 15.2 mmol/L transforms 1 mmol of substrate per minute, in standard conditions.
AMP 25 mmol/L The concentration is expressed in units per liter of sample (U/L).
P1,P5-di (adenosine-5’-) penta-phosphate 103 mmol/L
Glucose-6-phosphate Dehydrogenase (G6PDH) 9 KU/L Expected values
Creatine phosphate 250 mmol/L
Hexokinase (HK) 3 KU/L Men 24 - 204 U/L
Women 24 - 173 U/L
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Quality Control
immediately with plenty of soap and water. In case of severe injuries; Normal & abnormal commercial control serum of known
seek medical advice immediately. concentrations should be analyzed with each run.

The reagents are stable up to the expiration date specified when
stored at 2 – 8 oC.
84 85
CREATINE KINASE MB SYMBOLS IN PRODUCT LABELLING
Calculation
References
1. IFCC methods for the measurement of catalytic concentration
(CK-MB) EC REP Authorised Representative
DA/min x 8254 = U/L CKMB
of enzymes. Part 7: IFCC method for creatine kinase. JIFCC
1989; 1: 130-139.
LOT Batch Code/Lot number CAUTION. Consult instructions Units: One international unit (IU) is the amount of enzyme that
REF: 239 001 (6 x 5 ml) 30 Test
Catalogue Number for use transforms 1 mmol of substrate per minute, in standard conditions. 2. Tietz Textbook of Clinical Chemistry, 3rd edition. Burtis CA,
REF: 239 002 (6 x 10 ml) 60 Test REF
Consult instructions for use Manufactured by The concentration is expressed in units per liter of sample (U/L). Ashwood ER. WB Saunders Co., 1999.
REF: 239 003 (5 x 25 ml) 125 Test
REF: 239 004 (6 x 20 ml) 120 Test 3. Friedman and Young. Effects of disease on clinical laboratory
Expected values
The discrimination value for myocardial infarction is around 25 tests, 3th ed. AACC Press, 1997.
Intended Use Precautions and Warnings U/L. However, an index higher than 6% of total CK concentration 4. Urdal P and Landaas S. Clin Chem 1979; 25: 461-465.
Spectrum Diagnostics Creatine Kinase MB (CK-MB) reagent is Do not ingest or inhalate. In case of contact with eyes or skin; rinse discriminates better. These values are for orientation purpose; each
intended for the in-vitro quantitative, diagnostic determination of immediately with plenty of soap and water. In case of severe injuries; 5. Young DS. Effects of drugs on clinical laboratory tests, 3th ed.
laboratory should establish its own reference range.
Creatine kinase MB in human serum on both automated and manual seek medical advice immediately. AACC Press, 1997.
systems.
Quality Control
Storage & Stability Normal & abnormal commercial control serum of known
Background All the components of the kit are stable until the expiration date on concentrations should be analyzed with each run. ORDERING INFORMATION
Creatine kinase (CK) is an enzyme which is contained in heart, brain the label when stored tightly closed at 2-8oC, protected from light
and skeletal muscles. Thus, an increase of circulating level of CK and contaminations prevented during their use. Signs of reagent CATALOG NO QUANTITY
may be associated to myocardial infarction, acute cerebrovascular Performance Characterstics
deterioration: Presence of particles and turbidity.
disease, trauma or diseases of skeletal muscles. After a myocardial Precision 239 001 6 x 5 ml
infarct, CK level begins raising between 4th and 6th hour after first 239 002 6 x 10 ml
Reagent preparation, Storage, and Stability Within run (Repeatiblity) 239 003 5 x 25 ml
acute symptoms, reaching the peak between 18th and 30th hour
REF: 239 001 add 1 ml from R2 to one bottle of R1; mix gently 239 004 6 x 20 ml
and coming back to normal values during the 3rd day. CK is present Level 1 Level 2
REF: 239 002 add 2 ml from R2 to one bottle of R1; mix gently
in three different isoenzymatic forms, which could be separated by
REF: 239 003 add 5 ml from R2 to one bottle of R1; mix gently n 20 20
electrophoresis or column chromatography; each form is originated
REF: 239 004 add 4 ml from R2 to one bottle of R1; mix gently
in different body tissues, paying off their diagnostic determinations. Mean (U/L) 45 129
Or prepare the working solution according to the number of test
CK exists in serum in dimeric forms as CK-MM, CK-MB, and CK-BB
required by mixing 4 volumes of R1 with 1 volume of R2. Stability: 2 CV% 3.5 3.2
and as macro-enzymes. Measurement of CK-MB is a quite specific
weeks at 2-8oC away from light sources.
test for detection of cardiac muscle damage and is therefore used for
diagnosis and monitoring of myocardial infarction.
Specimen Collection and Preservation Run to run (Reproducibility)
Serum free of hemolysis is the preferred specimen. Plasma
Method containing heparin, EDTA, citrate or fluoride may produce
Level 1 Level 2
After immunoinhibition with antibodies to the CK-M subunit, the
unpredictable reaction rates. Stable for 2 hours at 20-25 oC, 5 days n 20 20
CK-B activity is determined with a method according to the
at 4-8 oC.Total CK concentration in the sample must be lower than
recommendations of the International Federation of Clinical Mean (U/L) 40 130
1000 U/L. Dilute the serum 1/2 if necessary, with NaCl (150 mmol/L).
Chemistry (IFCC). CV% 2.8 2.3
System Parameters
Assay Principle Wavelength 340 nm (334-365 nm) Methods Comparison
A specific antibody inhibits the M subunits of CK-MM and CK-MB, A comparison between Spectrum Diagnostics CK-MB reagent and a
Optical path 1 cm
and thus allows determination of the B subunit of CK-MB (assuming commercial reagent of the same methodology was performed on 20
Assay type Kinetic
the absence of CK-BB or CK-1). CK-B catalytic concentration, human sera. A correlation of 0.959 was obtained.
Direction Increase
which corresponds to half of CK-MB concentration, is determined
from the rate of NADPH formation, measured at 340 nm, by means
e.g.: Reagent volume 1 ml Sensitivity
of the hexokinase (HK) and glucose-6-phosphate dehydrogenase
Sample volume 40 ml When run as recommended, the minimum detection limit of the
(G6PDH) coupled reactions1,3. o
Temperature 37 C assay is 2.0 U/L.
CK Equilibration Time 60 seconds
Creatine phosphate + ADP Creatine + ATP
Read time 1 to 5 minutes Linearity
HK Zero adjustment against air The reaction is linear up to CK-MB concentration of 2000 U/l;
ATP + Glucose ADP + Glucose-6-phosphate
Reagent blank limits specimens showing higher concentration should be diluted 1+2
+ G6PDH + Sensitivity 2 U/L using physiological saline and repeat the assay (result×3).
Glucose-6- phosphate+NADP 6-Phosphogluconate+NADPH + H
Linearity 2000 U/L
Reagents Interferences:
Reagent 1 (pH 6.7) (Buffer / Coenzyme)
Procedure Haemoglobin (< 2.5 g/L) ,lipemia (Lipids < 900 mg/dL) and Bilirubin
1. Pipette into a thermostatized cuvette: (< 25 mg/dL) do not interfere. Presences in the sample of above
Imidazol 125 mmol/L normal concentrations of CK-BB or adenilate kinase, and of macro
D-Glucose 25 mmol/L Working solution 1 ml
or mitochondrial CK interfere. Other drugs and substances may
N-Acetyl-L-Cysteine 25 mmol/L Serum 40 mL interfere3,4.
Magnesium acetate 12.5 mmol/L
NADP 2.5 mmol/L
EDTA 2 mmol/L 2. Mix and incubate 60 seconds.
3. Read initial absorbance (A) of the sample, start the stopwatch
and read absorbance at 1 minute intervals thereafter for 5
Reagent 2 (Enzymes) minutes.
ADP 15.2 mmol/L 4. Calculate the difference between absorbances and the
AMP 25 mmol/L average absorbance differences per minute (DA/min).
P1,P5-di (adenosine-5’-) penta-phosphate 103 mmol/L
Glucose-6-phosphate Dehydrogenase (G6PDH) 9 KU/L
Creatine phosphate 250 mmol/L
Hexokinase (HK) 3 KU/L
Anti-human-CK-M.
86 87
g - Glutamyltransferase- SYMBOLS IN PRODUCT LABELLING
Precision
Analytical Range
2 – 600 U/L.
(gGT)-Liquizyme (9+1) EC REP Authorised Representative
Use by/Expiration Date Within run (Repeatiblity) Waste Disposal
E.C.2.3.2.2. LOT
REF
Catalogue Number
for use
Level 1 Level 2
REF: 246 001 ( 4 x 20 ml) 80 test Consult instructions for use Manufactured by n 20 20 S56: dispose of this material and its container at hazardous or
REF: 246 002 (10 x 10 ml) 100 test special waste collection point.
Mean (U/L) 44.75 120.2
REF: 246 003 ( 9 x 20 ml) 180 test S57: use appropriate container to avoid environmental
SD 2.07 2.2 contamination.
REF: 246 004 ( 4 x 60 ml) 240 test
REF: 246 005 ( 5 x 20 ml) 100 test CV% 4.63 1.84 S61: avoid release in environment. refer to special instructions/
Precautions and Warnings safety data sheets.
Intended Use immediately with plenty of soap and water. In case of severe injuries;
Spectrum Diagnostics liquizyme g-glutamyltransferase reagent is Run to run (Reproducibility) References
intended for the in-vitro quantitative, diagnostic determination of Level 1 Level 2 1. Heersink W, Hafkenscheid JCM, Siepel H, van der
g-glutamyltransferase in human serum on both automated and Both reagents (R1) and (R2) contain sodium azide which may react venjongekryg J, Dijt CCM. Temperature – converting factors
manual systems. n 20 20
with copper or lead plumbing. for enzymes: comparison of methods. Enzyme. 1980;25:333-
Mean (U/L) 45.1 121.3 341.
Background Reagent Storage and Stability SD 2.19 2.29
g- Glutamyltransferase (gGT) is usually most significantly elevated 2. Moss DW, Henderson AR, Kachmar IF. Enzymes In:Tietz NW,
by obstructive disease and has good specificity for the liver. It is o CV% 4.72 1.92 ed. Fundamentals of clinical chemistry. 3 rd ed.
stored refrigerated at 2 - 8 C. Working solution is stable for 3 months
not elevated in bone diseases or pregnancy (as ALP) or in skeletal o o
at 2 – 8 C or 2 weeks at 15 to 25 C when stored in a dark bottle. 3. Persjn JP, van der slike W. A new method for the determination
muscle diseases (as AST). gGT can also help to differentiate Methods Comparison of g-glutamyl transferase in serum. J Clin Chem Clin Biochem.
between mechanical and viral from drug induced cholestasis. The Deterioration A comparison between Spectrum Diagnostics g-GT and a commercial 1976;14421-427.
highest concentration of gGT is found in the luminal membrane of Do not use liquizyme gGT reagent if it is turbid or if the absorbance of reagent of the same methodology was performed on 20 human sera.
the proximal tubules of the kidney. Other sources are the pancreas, A correlation of 0.969 was obtained. 4. Saw M, Stromme JH, london JL, Theodorsen L. IFCC method
the working reagent is greater than 1.0 at 405 nm. Failure to recover
prostate, and liver. High gGT activity is found in prostate tissue, for g-glutamyl transferase[(g-glutamyl ) – peptide:ammino
which may account for the increased gGT activity seen in some sera Sensitivity acid g-glutamyl transferase, EC 2.3.2.2]. Clin Chem Acta. 1983;
from men compared with sera from women. When run as recommended, the minimum detection limit of this 135:315F-338F.
Specimen Collection and Preservation assay is 2.0 U/L. 5. Szasz, G., Persijn JP. Clin. Chem. Clin. Biochem. 1974;12:228.
Method Use serum and plasma, free from haemolysis. Heparin is the only
Kinetic colorimetric according to Szasz method.
(5)
acceptable anticoagulant. The biological half-life of gGT in serum is Linearity
3 – 4 days. The reaction is linear up to g-Glutamyltransferase concentration of
Assay Principle 600 U/L; specimens showing higher concentration should be diluted ORDERING INFORMATION
Stability:
o
7 days at 4 – 8 C ; 7 days at 20 – 25 C ;
o
1+5 with physiological saline and repeat the assay (result×6). CATALOG NO QUANTITY
Determination of g-Glutamyltransferase (g-GT) according to the o
1 year at -20 C
following reaction: 246 001 4 x 20 ml
246 002 10 x 10 ml
System Parameters Serum, plasma 246 003 9 x 20 ml
L-g-Glutamyl-3-carboxy-4-nitroanilide + Glycylglycine
Wavelength 405 nm (400 – 420 nm) 246 004 4 x 60 ml

Optical path 1 cm Haemolysis 246 005 5 x 20 ml
g-GT
Assay type Kinetic No significant interference up to a haemoglobin level of 5 g/L.

Direction Increase
L-g-Glutamyl- glycylglycine + 5-amino-2-nitrobenzoate
Sample : Reagent Ratio 1 : 10 Icterus
e.g.: Reagent volume 1 ml No significant interference.
The rate of liberation of yellow coloured indicator 5-amino-2-
Sample volume 100 ml
nitrobenzoate is directly proportional to g-GT activity in the sample o o
Temperature 37 C or 30 C Lipemia
and is quantitated by measuring the increase in absorbance at
Equilibration time 30 seconds. Lipemic specimens may cause high absorbance flagging.
405nm.
Read time 1 to 3 minutes Diluted sample treatment may be recommended.
Zero adjustment Against air
Reagents Reagent Blank Limits Low 0.2 AU Anticoagulants
High 1.0 AU Citrate, EDTA and fluoride inhibit the enzyme activity.
Reagent 1 (R1 Buffer) Sensitivity 2.0 U/L
Tris buffer pH 8.2 80 mmol/L Linearity 600 U/L Expected Values
Glycylglycine 130 mmol/L
Sodium Azide 8.0 mmol/L o
Procedure 37 C Females 7 -32 U/L (0.12 -0.53 mkat/L)
Males 11-50 U/L (0.18 -0. 82 mkat/L)
Reagent 2 (R2 Starter) Pipette in a test tube: Macro Semi-Micro
L-g-Glutamyl-3-carboxy-4-nitroanilide 4.0 mmol/L Working 1.0 ml 500 ml
Sodium Azide 8.0 mmol/L Solution o
30 C Females 5-24 U/L (0.08-0. 4 mkat/L)
Specimen 100 ml 50 ml Males 8-37 U/L (0.1 -0. 6 mkat/L)
For further information, refer to the g-Glutamyltrasferase reagent
material safety data sheet.
Mix, read initial absorbance after 30 seconds and start timer o
Reagent Preparation simultaneously. Read again after 1, 2 and 3 minutes. Determine the
Males 6-28 U/L (0. 1-0. 5 mkat/L)
Prepare working solution as following: mean absorbance change per minute (DA/min).
REF: 246 001: add 2 ml from R2 to one bottle of R; mix gently. Calculation
REF: 246 002: add 1 ml from R2 to one bottle of R; mix gently. To calculate the g-glutamyl transferase (g-GT) activity, use the Spectrum Diagnostics does not interpret the results of a
REF: 246 003: add 2 ml from R2 to one bottle of R; mix gently. following formula: clinical laboratory procedure; interpretation of the results is
REF: 246 004: add 6 ml from R2 to one bottle of R; mix gently. U/L = 1158 × DA 405 nm/min considered the responsibility of qualified medical personnel.
REF: 246 005: add 2 ml from R2 to one bottle of R; mix gently. All indications of clinical significance are supported by
Or prepare the working solution according to the number of tests Quality Control literature references.
required by mixing 9 volumes of reagent 1 (R1) and 1volume of Normal & abnormal control serum of known concentrations should
reagent 2 (R2) ,e.g. 900 ml R1 +100 ml R2. be analyzed with each run.
88 89
g - Glutamyltransferase- SYMBOLS IN PRODUCT LABELLING
Level 1 Level 2 1. Heersink W, Hafkenscheid JCM, Siepel H, van der
(gGT)-Liquizyme (1+1) EC REP Authorised Representative
Use by/Expiration Date n 20 20 venjongekryg J, Dijt CCM. Temperature – converting factors
for enzymes: comparison of methods. Enzyme. 1980;25: 333-
E.C.2.3.2.2. LOT
REF
Catalogue Number
for use
Mean (U/L) 45.1 121.3 341.
SD 2.19 2.29
REF: 247 001 (2 x 25 ml) 50 test Consult instructions for use Manufactured by 2. Moss DW, Henderson AR, Kachmar IF. Enzymes In :Tietz
REF: 247 002 (4 x 25 ml) 100 test CV% 4.72 1.92 NW, ed. Fundamentals of clinical chemistry. 3 rd ed.
3. Persjn JP, van der slike W. A new method for the determination
Intended Use Methods Comparison
Deterioration A comparison between Spectrum Diagnostics g-GT reagent and a
of g-glutamyl transferase in serum. J Clin Chem Clin Biochem.
Spectrum Diagnostics liquizyme g- glutamyltransferase reagent is Do not use liquizyme gGT reagent if it is turbid or if the absorbance of 1976;14421-427.
intended for the in-vitro quantitative, diagnostic determination of commercial reagent of the same methodology was performed on 20
the working reagent is greater than 1.0 at 405 nm. Failure to recover
g-glutamyltransferase in human serum on both automated and human sera. A correlation of 0.969 was obtained. 4. Saw M, Stromme JH, london JL, Theodorsen L. IFCC method
manual systems. for g-glutamyl transferase[(g-glutamyl ) – peptide:ammino
Sensitivity acid g- glutamyl transferase, EC 2.3.2.2]. Clin Chem Acta.
When run as recommended, the minimum detection limit of this 1983; 135:315F-338F.
Background Specimen Collection and Preservation
g-Glutamyltransferase (gGT) is usually most significantly elevated by assay is 2.0 U/L.
Use serum and plasma, free from hemolysis. Heparin is the only 5. Szasz, G., Persijn JP. Clin. Chem. Clin. Biochem. 1974;12:228.
obstructive disease and has good specificity for the liver. It is not acceptable anticoagulant. The biological half-life of gGT in serum is
elevated in bone diseases or pregnancy (as in ALP) or in skeletal Linearity
3 – 4 days.
muscle diseases (as AST). gGT can also help to differentiate o
Stability: 7 days at 4 – 8 C ; 7 days at 20 – 25 C ;
o
between mechanical and viral from drug induced cholestasis. The o The reaction is linear up to g-Glutamyltransferase concentration of ORDERING INFORMATION
1 year at -20 C
highest concentration of gGT is found in the luminal membrane of 600 U/L; specimens showing higher concentration should be diluted
1+5 with physiological saline and repeat the assay (result×6). CATALOG NO QUANTITY
the proximal tubules of the kidney. Other sources are the pancreas, System Parameters
prostate, and liver. High gGT activity is found in prostate tissue, 247 001 2 x 25 ml
which may account for the increased gGT activity seen in some sera Interfering Substances 247 002 4 x 25 ml
Optical path 1 cm
from men compared with sera from women. Serum, plasma
Assay type Kinetic
Direction Increase
Method Hemolysis
Kinetic colorimetric according to Szasz method.
(5) No significant interference up to a hemoglobin level of 5 g/L.
Icterus
Assay Principle Temperature
o
37 C or 30 C
o
No significant interference.
Determination of g-Glutamyltransferase (gGT) according to the Equilibration time 30 seconds.
following reaction: Read time 1 to 3 minutes
Lipemia
Lipemic specimens may cause high absorbance flagging
L-g-Glutamyl-3-carboxy-4-nitroanilide + Glycylglycine Reagent Blank Limits Low 0.2 AU
Diluted sample treatment may be recommended.
High 1.0 AU
g-GT Sensitivity 2.0 U/L
Anticoagulants
Linearity 600 U/L
Citrate, EDTA and fluoride inhibit the enzyme activity.
L-g-Glutamyl- glycylglycine + 5-amino-2-nitrobenzoate
Procedure Expected Values
The rate of liberation of yellow coloured indicator 5-amino-2-
nitrobenzoate is directly proportional to g-GT activity in the sample Pipette in a test tube: o
37 C Females 7 -32 U/L (0.12 -0.53 mkat/L)
and is quantitated by measuring the increase in absorbance at working solution 1.0 ml (or add 0.5 ml R1 + 0.5 ml R2) Males 11-50 U/L (0.18 -0. 82 mkat/L)
405nm. o
Specimen 100 ml 30 C Females 5-24 U/L (0.08-0. 4 mkat/L)
Males 8-37 U/L (0.1 -0. 6 mkat/L)
Reagents Mix, read initial absorbance after 30 sec. and start timer simult- o
aneously. Read again after 1, 2 and 3 minutes. Determine the mean Males 6-28 U/L (0. 1-0. 5 mkat/L)
Reagent 1 (R1 Buffer)
absorbance change per minute (DA/min).
Tris buffer pH 8.2 120 mmol/L
Glycylglycine 300 mmol/L
Sodium Azide 12 mmol/L
Calculation
To calculate the g-glutamyl transferase (gGT) activity, use the
following formula:
Reagent 2 (R2 Starter) considered the responsibility of qualified medical personnel.
U/L = 1158 × DA 405 nm /min
L-g-Glutamyl-3-carboxy-4-nitroanilide 1.0 mmol/L All indications of clinical significance are supported by
Sodium Azide 8 mmol/L literature references.
Quality Control
For further information, refer to the g-Glutamyltrasferase reagent Normal & abnormal control serum of known concentrations should
material safety data sheet. be analyzed with each run.
Analytical Range
Precautions and Warnings Performance Characterstics 2 – 600 U/L.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Precision
immediately with plenty of soap and water. In case of severe injuries; Waste Disposal
seek medical advice immediately. Within run (Repeatiblity) This product is made to be used in professional laboratories.
Level 1 Level 2
Both reagents (R1) and (R2) contain sodium azide which may react S56: dispose of this material and its container at hazardous or
with copper or lead plumbing. n 20 20 special waste collection point.
Mean (U/L) 44.75 120.2 S57: use appropriate container to avoid environmental
Reagent Preparation, Storage and Stability contamination.
All reagents are stable until expiration date stated on label when SD 2.07 2.2 S61: avoid release in environment. refer to special instructions/
o
stored refrigerated at 2 - 8 C. CV% 4.63 1.84 safety data sheets.
Working solution can be prepared by adding equall volumes from
o o
R1 and R2 ; Stability : 3 months at 2 - 8 C or 2 weeks at 15 -25 C
when stored in a dark bottle.
90 91
Aspartate aminotransferase SYMBOLS IN PRODUCT LABELLING Procedure Drugs
Calcium dobesilate and doxycycline HCL cause artificially low
(AST/GOT)-Liquizyme (4+1) EC REP Authorised Representative
CAUTION. Consult instructions Working
Macro
1.0 ml
Semi-Micro
500 ml
AST values at the tested drug level.
E.C.2.6.1.1. LOT Batch Code/Lot number

Catalogue Number
for use
Manufactured by
solution Expected values
REF Specimen 100 ml 50 ml o
37 C Females up to 31 U/l (up to 0.52 mkat/L)
REF: 291 001 ( 4 x 20 ml) 80 test Consult instructions for use (Xi) - Irritant
Males up to 37 U/l (up to 0.62 mkat/L)
REF: 291 002 (10 x 10 ml) 100 test Temperature Limitation Mix, read initial absorbance after 60 seconds. and start timer o
REF: 291 003 ( 9 x 20 ml) 180 test 30 C Females up to 21 U/l (up to 0.35 mkat/L)
simultaneously. Read again after 1, 2 and 3 minutes.
REF: 291 004 ( 4 x 60 ml) 240 test Males up to 25 U/l (up to 0.42 mkat/L)
Irritant (Xi): R36/38: Irritating to eyes and skin. S26: In case of Determine the mean absorbance change per minute (DA/min).
REF: 291 005 ( 5 x 20 ml) 100 test Temperature conversion factor is 1.37 (25
o
30 C ) and
contact with eyes, rinse immediately with plenty of water and
REF: 291 006 ( 4 x 50 ml) 200 test Calculation 2.04 (25
o
37 C ).
seek medical advice. S37/39: Wear suitable gloves and eye/face
REF: 291 007 ( 5 x100 ml) 500 test To calculate the AST/GOT activity use the following formulae:
protection.
REF: 291 008 ( 6 x100 ml) 600 test U/l = 1780 x DA 334 nm /min
Reagent 2 (R2 Coenzyme) U/l = 1746 x DA 340 nm /min Spectrum Diagnostics does not interpret the results of a
Intended Use NADH ≥ 0.18 mmol/L U/l = 3235 x DA 365 nm /min clinical laboratory procedure; interpretation of the results is
Spectrum Diagnostics liquizyme AST reagent is intended for the in- 2 – Oxoglutarate 18 mmol/L considered the responsibility of qualified medical personnel.
vitro quantitative, diagnostic determination of AST in human serum Sodium Azide 8 mmol/L Quality Control All indications of clinical significance are supported by
on both automated and manual systems. For further information, refer to the Aspartate aminotransferase Normal & abnormal control serum of known concentrations should literature references.
reagent material safety data sheet. be analyzed with each run.
Background
The enzyme aspartate aminotransferase (AST) is widely distributed
Analytical Range
Precautions and Warnings Performance Characterstics 5 – 400 U/L.
in erythrocytes and tissues, principally heart, liver, muscle, and Do not ingest or inhalate. In case of contact with eyes or skin; rinse Precision
kidney. Elevated serum levels are found in diseases involving these immediately with plenty of soap and water. In case of severe injuries;
tissues such as myocardial infarction,viral hepatitis and muscular Waste Disposal
seek medical advice immediately. Within run (Repeatiblity) This product is made to be used in professional laboratories.
dystrophy. Following myocardial infarction, serum AST is elevated
and reaches a peak two days after onset. Two isoenzymes of Level 1 Level 2 Please consult local regulations for a correct waste disposal.
Both reagents (R1) and (R2) contains sodium azide which may react S56: dispose of this material and its container at hazardous or
AST have been detected, cytoplasmic and mitochondrial. Only with copper or lead plumbing. n 20 20 special waste collection point.
the cytoplasmic isoenzyme occurs in normal serum, while the
mitochondrial, together with the cytoplasmic isoenzyme, has been Mean (U/L) 32.6 133 S57: use appropriate container to avoid environmental
Reagent Preparation contamination.
detected in the sera of patients with coronary and hepatobiliary SD 1.3 1.3
Prepare working solution as following: S61: avoid release in environment. refer to special instructions/
diseases.
REF:291 001 : add 4 ml from R2 to one bottle of R1; mix gently. CV% 4.08 0.97 safety data sheets.
REF:291 002 : add 2 ml from R2 to one bottle of R1; mix gently.
Method REF:291 003 : add 4 ml from R2 to one bottle of R1; mix gently.
Kinetic method according to the International Federation of References
REF:291 004 : add 12 ml from R2 to one bottle of R1; mix gently. Run to run (Reproducibility)
ClinicalChemistry (IFCC) (3). REF:291 005 : add 4 ml from R2 to one bottle of R1; mix gently. 1. Breuer J, Report on the symposium “drug effects in
REF:291 006 : add 10 ml from R2 to one bottle of R1; mix gently. Level 1 Level 2 clinicalchemistry methods”. Eur J Clin Chem Clin Biochem.
Assay Principle REF:291 007 : add one bottle of R2 to one bottle of R1; mix gently. n 20 20 1996;34:385-386.
The series of the reaction involved in the assay system is as follows: REF:291 008 : add one bottle of R2 to one bottle of R1; mix gently.
Or prepare the working solution according to the number of tests Mean (U/L) 33.1 135.5 2. ECCLS. Determination of the catalytic activity concentration in
1. The amino group is enzymatically transferred by AST required by mixing 4 volumes of reagent 1 (R1) and 1volume of SD 1.5 1.42 serum on L- aspartate aminotransferase (EC 2.6.1.1,AST) Clin
present in the sample from L-aspartate to the carbon atom of reagent 2 (R2), e.g. 400 ml R1 +100 ml R2. Chem. 1989;20:204-211.
2-oxoglutarate yielding oxaloacetate and L-glutamate. CV% 4.25 1.13
3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
Oxaloacetate Reagent Storage and Stability
L-Aspartate AST All reagents are stable until expiration date stated on label when
Methods Comparison Biochem 1986;24:481-95.
+ + o A comparison between Spectrum Diagnostics AST (4+1) reagent
2-Oxoglutarate L-Glutamate stored refrigerated at 2 - 8 C. Working solution is stable for 4 weeks
o o and a commercial reagent of the same methodology was performed 4. Henry RJ, et al. Am j Clin Path 1960 :34:381
at 2 – 8 C or 2 days at 15 - 25 C.
on 20 human serum.A correlation of 0.991 was obtained.
5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ,
Deterioration eds.Clinical chemistry, theory, analysis, and correlation.
2. Oxaloacetate in presence of NADH and malate dehyrogenase Do not use liquizyme AST reagent if it is turbid or if the absorbance
Sensitivity
When run as recommended, the minimum detection limit of this Stlouis:mosby;1984:420- 438.
(MDH), is reduced to L-malate. In this reaction NADH is of the working reagent is less than 1.0 at 340 nm. Failure to recover
oxidized to NAD. The reaction is monitored by measuring the assay is 5.0 U/L.
control values within the assigned range may be an indication of 6. Young DS. Effects of drugs on clinical laboratory tests.Third
rate of decrease in absorbance at 340 nm due to oxidation of reagent deterioration. edition. 1990 :3:6-12.
NADH to NAD. Linearity
Specimen Collection and Preservation The reaction is linear up to AST concentration of 400 U/L; 7. Zilva JF, pannall PR : Plasma enzymes in diagnosis inclinical
Oxaloacetate L-Lactate specimens showing higher concentration should be diluted 1+5 with
MDH + Use nonhemolyzed serum. Heparin and EDTA are the only chemistry in diagnosis and treatment lioydluke london
+ physiological saline and repeat the assay (result×6).
NAD+ acceptable anticoagulants. The biological half-life of AST in serum 1979:chap 17: 338.
NADH + H+
is 17 hours.
o
Stability: 1 day at 15 – 25 C; 7 days at 4 - 8 C;
o
o
12 weeks at -20 C Serum, plasma ORDERING INFORMATION
3. Addition of lactate dehydrogenase (LDH) to the reagent
is necessary to achieve rapid and complete reduction of CATALOG NO QUANTITY
System Parameters Hemolysis
endogenous pyruvate so that it does not interfere with the Erythrocyte contamination elevates results, since AST activities 291 001 4 x 20 ml
assay. in erythrocytes are 15 times higher than those in normal sera. 291 002 10 x 10 ml
Optical path 1 cm
291 003 9 x 20 ml
L-Lactate Assay type Kinetic
Sample pyruvate LDH Icterus 291 004 4 x 60 ml
+ + Direction decrease
No significant interference. 291 005 5 x 20 ml
NADH + H+ NAD+ Sample : Reagent Ratio 1 : 10
291 006 4 x 50 ml
Lipemia 291 007 5 x 100 ml
Reagents o o Lipemic specimens may cause high absorbance flagging. 291 008 6 x 100 ml
Temperature 37 C or 30 C
Reagent 1 (R1 Buffer / Enzymes) Equilibration time 60 seconds. Diluted sample is recommended.
Tris buffer (pH 7.7 ) 80 mmol/L Read time 1 to 3 minutes
L- Aspartate 240 mmol/L Zero adjustment Against air Anticoagulants
MDH ≥ 450 U/L Reagent Blank Limits Low 1.00 AU Citrate and fluoride inhibit the enzyme activity.
LDH ≥ 1200 U/L High 2.5 AU
Sodium Hydroxide 220 mmol/L Sensitivity 5 U/L
Sodium Azide 8 mmol/L Linearity 400 U/L
92 93
Aspartate aminotransferase SYMBOLS IN PRODUCT LABELLING
Calculation
To calculate the AST/GOT activity use the following formulae
Expected values
(AST/GOT)-Liquizyme (1 + 1)
o
EC REP Authorised Representative Use by/Expiration Date U/l = 1780 x DA 334 nm /min 37 C Females up to 31 U/l (up to 0.52 mKat/L)
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions U/l = 1746 x DA 340 nm /min Males up to 37 U/l (up to 0.62 mKat/L)
E.C.2.6.1.1. LOT Batch Code/Lot number
Catalogue Number
for use
Manufactured by
U/l = 3235 x DA 365 nm /min
o
30 C Females up to 21 U/l (up to 0.35 mKat/L)
REF
Consult instructions for use (Xi) - Irritant Quality Control Males up to 25 U/l (up to 0.42 mKat/L)
REF: 259 001 (2 x 25 ml) 50 test
o
concentrations should be analyzed with each run. Temperature conversion factor is 1.37 ( 25 30 C )
REF: 259 003 (2 x 100 ml) 200 test o
and 2.04 ( 25 37 C ).
Reagent 2 (R2 Coenzyme) Performance Characteristics
Intended Use NADH ≥ 0.06 mmol/L
Spectrum Diagnostics liquizyme AST reagent is intended for the in- Precision Spectrum Diagnostics does not interpret the results of a
2-Oxoglutarate 4 mmol/L Within run (Repeatiblity) clinical laboratory procedure; interpretation of the results is
vitro quantitative, diagnostic determination of AST in human serum Sodium Azide 8 mmol/L
on both automated and manual systems. considered the responsibility of qualified medical personnel.
Level 1 Level 2
For further information, refer to the Aspartate aminotransferase n 20 20 literature references.
Background reagent material safety data sheet.
The enzyme aspartate aminotransferase (AST) is widely distributed Mean (U/L) 32 135
in erythrocytes and tissues, principally heart, liver, muscle, and Precautions and Warnings SD 1.2 1.27 Analytical Range
kidney. Elevated serum levels are found in diseases involving these Do not ingest or inhalate. In case of contact with eyes or skin; rinse 5 – 400 U/L.
tissues such as myocardial infarction, viral hepatitis and muscular CV% 4.1 0.99
dystrophy. Following myocardial infarction, serum AST is elevated seek medical advice immediately.
Run to run (Reproducibility) Waste Disposal
and reaches a peak two days after onset. Two isoenzymes of
AST have been detected, cytoplasmic and mitochondrial. Only Reagent (R1) contains sodium azide which may react with copper Level 1 Level 2 Please consult local regulations for a correct waste disposal.
the cytoplasmic isoenzyme occurs in normal serum, while the or lead plumbing. n 20 20 S56: dispose of this material and its container at hazardous or
mitochondrial, together with the cytoplasmic isoenzyme, has been
detected in the sera of patients with coronary and hepatobiliary Mean (U/L) 34 136
Reagent Preparation, Storage and Stability S57: use appropriate container to avoid environmental
diseases.
All reagents are stable until expiration date stated on label when SD 1.6 1.43 contamination.
o
stored refrigerated at 2 - 8 C. S61: avoid release in environment. refer to special instructions/
Method Working solution can be prepared by adding equal volumes from R1
CV% 4.5 1.16
safety data sheets.
Kinetic method according to the International Federation of Clinical o
and R2; Stability: 2 days at 2 – 8 C.
Chemistry (IFCC) (3). Methods Comparison References
A comparison between Spectrum Diagnostics AST (1+1) reagent
Deterioration
Assay Principle Do not use liquizyme AST reagent if it is turbid or if the absorbance
and a commercial reagent of the same methodology was performed 1. Breuer J, report on the symposium “drug effects in clinical
The series of the reaction involved in the assay system is as follows: on 20 human sera. A correlation of 0.98 was obtained. chemistry methods”. Eur J Clin Chem clin Biochem.
of the working reagent is less than 1.0 at 340 nm.
Failure to recover control values within the assigned range may be 1996;34:385-386.
1. The amino group is enzymatically transferred by AST an indication of reagent deterioration. Sensitivity
2. ECCLS. Determination of the catalytic activity concentration
present in the sample from L-aspartate to the carbon atom of When run as recommended, the minimum detection limit of this
in serum on L- aspartate aminotransferase (EC 2.6.1.1,AST)
2-oxoglutarate yielding oxaloacetate and L-glutamate. assay is 5.0 U/L.
Specimen Collection and Preservation Clin Chem. 1989;20:204-211.
Oxaloacetate Use nonhemolyzed serum. Heparin and EDTA are the only
L-Aspartate AST Linearity 3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
+ + acceptable anticoagulants. The biological half-life of AST in serum
is 17 hours. The reaction is linear up to AST concentration of 400 U/L;specimens Biochem 1986;24:481-95.
2-Oxoglutarate L-Glutamate
o
Stability: 1 day at 15 – 25 C; 7 days at 4 - 8 C;
o showing higher concentration should be diluted 1+5 with physiological
4. Henry RJ et al. Am J Clin Bath 1960 :34:381
12 weeks at -20 C
o saline and repeat the assay (result×6).
2. Oxaloacetate in presence of NADH and malate dehydrogenase 5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ, eds.
(MDH), is reduced to L-malate. In this reaction NADH is Interfering Substances Clinical chemistry, theory,analysis, and correlation. St
System Parameters
oxidized to NAD. The reaction is monitored by measuring the Serum, plasma louis:mosby;1984:420- 438.
rate of decrease in absorbance at 340 nm due to oxidation of
Optical path 1 cm 6. Young DS. Effects of drugs on clinical laboratory tests. Third
NADH to NAD. Haemolysis
Assay type Kinetic edition. 1990 :3:6-12.
Direction decrease Erythrocyte contamination elevate results, since AST activities in
Oxaloacetate L-Lactate erythrocytes are 15 times higher than those in normal sera.
MDH + Sample: Reagent Ratio 1 : 10 7. Zilva JF, pannall PR : Plasma enzymes in diagnosis in
+
NAD+ e.g.: Reagent volume 1 ml clinical chemistry in diagnosis and treatment lioydluke london
NADH + H+ Icterus
Sample volume 100 ml 1979:chap 17 : 338.
o o No significant interference.
3. Addition of lactate dehydrogenase (LDH) to the reagent Equilibration time 30 seconds.
is necessary to achieve rapid and complete reduction of Read time 1 to 3 minutes Lipemia
endogenous pyruvate so that it does not interfere with the Lipemic specimens may cause high absorbance flagging. Diluted ORDERING INFORMATION
assay. Reagent Blank Limits Low 1.00 AU sample is recommended. CATALOG NO QUANTITY
High 2.5 AU
Sample pyruvate L-Lactate Anticoagulants 259 001 2 x 25 ml
LDH + Sensitivity 5 U/L
+ Citrate and fluoride inhibit the enzyme activity. 259 002 4 x 25 ml
NAD+ Linearity 400 U/L
NADH + H+ 259 003 2 x 100 ml
Drugs
Procedure
Reagents Calcium dobesilate and doxycycline HCL cause artificially low AST
Pipette in a test tube: values at the tested drug level.
Reagent 1 (R1 Buffer/Enzymes)
Tris buffer (pH 7.7) 80 mmol/L Working 1.0 ml ( or 0.5 ml R1 + 0.5 ml R2)
L- Aspartate 450 mmol/L solution
MDH ≥ 900 U/L
LDH ≥ 2000 U/L Specimen 100 ml
Sodium Hydroxide 300 mmol/L
Sodium Azide 8 mmol/L Mix, read initial absorbance after 30 seconds. and start timer
Irritant (Xi): R36/38: Irritating to eyes and skin. S26: In case of simultaneously. Read again after 1, 2 and 3 minutes. Determine
contact with eyes, rinse immediately with plenty of water and the mean absorbance change per minute (DA/min).
seek medical advice. S37/39: Wear suitable gloves and eye/face
protection.
94 95
Calculation References
1. Henry RJ et al. Am J Clin Path 1960 :34:381.
(AST/GOT)-Colorimetric EC REP Authorised Representative
Obtain the AST activity from the following table
2. Reitman S and Frankel S.Am .J.Clin.Path, 1975 ;28;65.
Absorbance U/L Absorbance U/L
LOT Batch Code/Lot number for use 3. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ, eds.
REF: 260 001 (2 x 50 ml) 100 test 0.020 7 0.100 36 Clinical chemistry, theory,analysis, and correlation. St
REF: 260 002 (2 x100 ml) 200 test REF Catalogue Number Manufactured by
0.030 10 0.110 41 louis:Mosby;1984:420- 438.
0.040 13 0.120 47 4. Young DS. Effects of drugs on clinical laboratory tests. Third
Intended Use Temperature Limitation (C) - Corrosive
0.050 16 0.130 52 edition. 1990 :3:6-12.
Spectrum Diagnostics colorimetric AST reagent is intended for 0.060 19 0.140 59
the in-vitro quantitative, diagnostic determination of AST in human 0.070 23 0.150 67
serum. Reagent (R1) contains sodium azide which may react with copper 0.080 27 0.160 76
or lead plumbing. 0.090 31 0.170 89 ORDERING INFORMATION
Background CATALOG NO QUANTITY
The enzyme aspartate aminotransferase (AST) is widely distributed Reagent preparation, Storage and Stability Quality Control
in erythrocytes and tissues, principally heart, liver, muscles, and 260 001 2 x 50 ml
The reagents are supplied ready-to-use and stable up to the expiry Normal & abnormal commercial control serum of known
kidneys. Elevated serum levels are found in diseases involving o 260 002 2 x 100 ml
date labeled on the bottles when stored at 2 – 8 C. concentrations should be analyzed with each run.
these tissues such as myocardial infarction, viral hepatitis and
muscular dystrophy. Following myocardial infarction, serum AST is Deterioration Sensitivity
elevated and reaches a peak two days after onset.Two isoenzymes Do not use The AST regents if precipitate forms. Failure to recover If run as recommended, the minimum detection level is 7 U/L.
of AST have been detected, cytoplasmic and mitochondrial. Only control values within the assigned range may be an indication of
the cytoplasmic isoenzyme occurs in normal serum, while the reagent deterioration. Linearity
mitochondrial, together with the cytoplasmic isoenzyme, has been
The assay is linear up to 89 U/L. If the absorbance exceeds 0.170
detected in the sera of patients with coronary and hepatobiliary
Specimen Collection and Preservation at 546 nm ( 89 U/L ), samples should be diluted 1 + 9 using sodium
diseases.
Use only non haemolyzed serum. The only acceptable anticoagulants chloride and repeat the assay (result × 10).
are heparin and EDTA. The biological half-life of AST in serum is 17
Method hours. Interfering Substances
AST – (Colorimetric method). o
Stability: 1 day at 15 – 25 C ; 7 days at 4 - 8 C ;
o
Serum, plasma
o
12 weeks at -20 C
Assay Principle Haemolysis
The reaction involved in the assay system is as follows: System Parameters Erythrocyte contamination elevates results, since AST activities
Wavelength 546 nm (530-550 nm) in erythrocytes are 15 times higher than those in normal sera.
The amino group is enzymatically transferred by AST present in Optical path 1 cm
the sample from L-aspartate to the carbon atom of 2-oxoglutarate Assay type Endpoint Icterus
yielding oxaloacetate and L-glutamate. Direction Increase No significant interference.
Oxaloacetate Sample : Reagent Ratio 1 : 60
L-Aspartate AST o o
+ + Temperature 37 C and 20 – 25 C Lipemia
2-Oxoglutarate L-Glutamate Zero adjustment Reagent or Sample blank Lipemic specimens may cause high absorbance flagging.
Sensitivity 7 U/L Diluted sample is recommended.
Linearity 89 U/L
Note
AST activity is measured by monitoring the concentration of
Procedure High concentration of aldehydes, ketones, or oxo-acids in some sera
oxaloacetate hydrazone formed with 2,4-dinitrophenylhydrazine.
1. Measurement against Reagent Blank may cause false high transaminases levels. Measurement aganist
a serum blank instead of a reagent blank avoids the risk of finding
Reagents such artifacts.
Pipette into test tubes
Phosphate buffer 100 mmol/L Reagent blank Sample
L- aspartate 100 mmol/L
Expected values
R1(buffer) 0.5 ml 0.5 ml Up to 12 U/L.
2–Oxoglutarate 5 mmol/L
Sample ------ 100 ml
Sodium Azide 12 mmol/L Distilled water 100 ml ------ Spectrum Diagnostics does not interpret the results of a
Harmful (Xn): R20/22: Harmful by inhalation and if swallowed. Mix and incubate for exactly 30 minutes at 37 C
o
S24/25: Avoid contact with skin and eyes. considered the responsibility of qualified medical personnel.
R2 0.5 ml 0.5 ml All indications of clinical significance are supported by
o
Reagent 2 (R2) Mix and incubate for exactly 20 minutes at 20 – 25 C literature references.
2,4-dinitrophenyl-hydrazine 2 mmo/L Sodium hydroxide 5.0 ml 5.0 ml
HCl 8.4 %
(C)-Corrosive contains caustic materials. Mix, measure absorbance of specimen against reagent blank at Analytical Range
R35 Causes severe burns. 546 nm after 5 minutes.a 7 – 89 U/L.
R41 Risk of serious damage to eyes.
S26 In case of contact with eyes, rinse immediately with 2. Measurement against Sample Blank Waste Disposal
plenty of water and seek medical advice. This product is made to be used in professional laboratories.
Sample blank Sample
S28 After contact with skin, wash immediately with plenty of Please consult local regulations for a correct waste disposal.
R1(buffer) 0.5 ml 0.5 ml S56: dispose of this material and its container at hazardous or
soap and water.
Sample ----- 100 ml special waste collection point.
For further information, refer to the Aspartate aminotransferase Mix and incubate for exactly 30 minutes at 37 C
o S57: use appropriate container to avoid environmental
reagent material safety data sheet. contamination.
R2 0.5 ml 0.5 ml S61: avoid release in environment. refer to special instructions/
Sample 100 ml ----- safety data sheets.
Additional Reagent o
Sodium hydroxide 0.4 mol/L. Mix and incubate for exactly 20 minutes. at 20 – 25 C
Sodium hydroxide 5.0 ml 5.0 ml
Mix, measure absorbance of specimen against sample
blank at 546 nm after 5 minutes.
96 97
Calculation
To calculate the AST/GOT activity use the following formulae:
Expected values
(AST/GOT) - Ultimate Single Reagent EC REP Authorised Representative Use by/Expiration Date
o
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions U/l = 1780 x DA 334 nm /min Males up to 37 U/l (up to 0.62 mKat/L)
E.C.2.6.1.1. t LOT Batch Code/Lot number for use U/l = 1746 x DA 340 nm /min
o
REF Catalogue Number Manufactured by U/l = 3235 x DA 365 nm /min 30 C Females up to 21 U/l (up to 0.35 mKat/L)
REF: 261 001 ( 2 x 20 ml) 40 test Consult instructions for use (Xi) - Irritant
Males up to 25 U/l (up to 0.42 mKat/L)
REF: 261 002 ( 6 x 20 ml) 120 test Temperature Limitation Quality Control o
REF: 261 003 ( 2 x 100 ml) 200 test Normal & abnormal control serum of known concentrations should Temperature conversion factor is 1.37 (25 30 C ) and 2.04
o
REF: 261 004 ( 4 x 50 ml) 200 test be analyzed with each run. (25 37 C ).
REF: 261 005 ( 2 x 50 ml) 100 test
Irritant (Xi): R36/38: Irritating to eyes and skin. S26: In case of Performance Characterstics Spectrum Diagnostics does not interpret the results of a
Intended Use contact with eyes, rinse immediately with plenty of water and
Precision clinical laboratory procedure; interpretation of the results is
Spectrum Diagnostics Ultimate AST reagent is intended for the in- seek medical advice. S37/39: Wear suitable gloves and eye/face considered the responsibility of qualified medical personnel.
vitro quantitative, diagnostic determination of AST in human serum protection. All indications of clinical significance are supported by
Level 1 Level 2
on both automated and manual systems. literature references.
The reagent also contains additives required to maintain NADH in n 20 20
its reduced forrm.
Background Mean (U/L) 32.6 133
The enzyme aspartate aminotransferase (AST) is widely distributed Analytical Range
For further information, refer to the •Aspartate aminotransferase SD 1.3 1.3 5 – 400 U/L.
in erythrocytes and tissues, principally heart, liver, muscle, and
reagent material safety data sheet.
kidney. Elevated serum levels are found in diseases involving these CV% 4.08 0.97
tissues such as myocardial infarction,viral hepatitis and muscular Waste Disposal
dystrophy. Following myocardial infarction, serum AST is elevated Precautions and Warnings This product is made to be used in professional laboratories.
and reaches a peak two days after onset. Two isoenzymes of Do not ingest or inhalate. In case of contact with eyes or skin; rinse Please consult local regulations for a correct waste disposal.
AST have been detected, cytoplasmic and mitochondrial. Only immediately with plenty of soap and water. In case of severe injuries; S56: dispose of this material and its container at hazardous or
the cytoplasmic isoenzyme occurs in normal serum, while the seek medical advice immediately. Level 1 Level 2 special waste collection point.
mitochondrial, together with the cytoplasmic isoenzyme, has been n 20 20 S57: use appropriate container to avoid environmental
detected in the sera of patients with coronary and hepatobiliary The Reagent (R) contain sodium azide which may react with copper contamination.
or lead plumbing. Mean (U/L) 33.1 135.5
diseases. S61: avoid release in environment. refer to special instructions/
SD 1.5 1.42 safety data sheets.
Method Reagent Preparation, Storage and Stability
Spectrum Ultimate AST reagent is supplied ready-to-use and stable CV% 4.25 1.13
Kinetic method according to the International Federation of References
ClinicalChemistry (IFCC)(3). up to the expiry date labeled on the bottles
Once opened, the opened vial is stable for 3 months at the specified Methods Comparison 1. Breuer J, Report on the symposium “drug effects in
temperature. A comparison between Spectrum Diagnostics AST reagent and a clinicalchemistry methods”. Eur J Clin Chem Clin Biochem.
Assay Principle commercial reagent of the same methodology was performed on 20 1996;34:385-386.
The series of the reaction involved in the assay system is as follows: human serum.A correlation of 0.991 was obtained.
Deterioration 2. ECCLS. Determination of the catalytic activity concentration in
1. The amino group is enzymatically transferred by AST Do not use Spectrum Ultimate AST reagent if it is turbid or if the
Sensitivity serum on L- aspartate aminotransferase (EC 2.6.1.1,AST) Clin
present in the sample from L-aspartate to the carbon atom of absorbance of the working reagent is less than 0.9 at 340 nm.
When run as recommended, the minimum detection limit of this Chem. 1989;20:204-211.
2-oxoglutarate yielding oxaloacetate and L-glutamate. Failure to recover control values within the assigned range may be
an indication of reagent deterioration. assay is 5.0 U/L. 3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
L-Aspartate Oxaloacetate Biochem 1986;24:481-95.
AST + Linearity
+ Specimen Collection and Preservation
L-Glutamate The reaction is linear up to AST concentration of 400 U/L; 4. Henry RJ, et al. Am j Clin Path 1960 :34:381
2-Oxoglutarate Use nonhemolyzed serum. Heparin and EDTA are the only
acceptable anticoagulants. The biological half-life of AST in serum specimens showing higher concentration should be diluted 1+5 with 5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ,
is 17 hours. physiological saline and repeat the assay (result×6). eds.Clinical chemistry, theory, analysis, and correlation.
2. Oxaloacetate in presence of NADH and malate dehyrogenase o o
(MDH), is reduced to L-malate. In this reaction NADH is Stability: 1 day at 15 – 25 C; 7 days at 4 - 8 C; Stlouis:mosby;1984:420- 438.
oxidized to NAD. The reaction is monitored by measuring the
o
12 weeks at -20 C Interfering Substances
Serum, plasma 6. Young DS. Effects of drugs on clinical laboratory tests.Third
rate of decrease in absorbance at 340 nm due to oxidation of
edition.1990 :3:6-12.
NADH to NAD. System Parameters
Wavelength 340 nm (334 – 365 nm) Hemolysis 7. Zilva JF, pannall PR : Plasma enzymes in diagnosis inclinical
Oxaloacetate L-Lactate Optical path 1 cm Erythrocyte contamination elevates results, since AST activities chemistry in diagnosis and treatment lioydluke london
MDH +
+ Assay type Kinetic in erythrocytes are 15 times higher than those in normal sera. 1979:chap 17: 338.
NADH + H+ NAD+ Direction decrease
Sample : Reagent Ratio 1 : 10 Icterus
3. Addition of lactate dehydrogenase (LDH) to the reagent e.g.: Reagent volume 1 ml No significant interference.
Sample volume 100 ml ORDERING INFORMATION
is necessary to achieve rapid and complete reduction of
endogenous pyruvate so that it does not interfere with the Temperature 37 C or 30 C
o o
Lipemia CATALOG NO QUANTITY
assay. Equilibration time 60 seconds. Lipemic specimens may cause high absorbance flagging.
261 001 2 x 20 ml
Read time 180 seconds Diluted sample is recommended.
L-Lactate 261 002 6 x 20 ml
Sample pyruvate LDH Zero adjustment Against air
+ + 261 003 2 x 100 ml
Reagent Blank Limits Low 0.9 AU Anticoagulants
NADH + H+ NAD+ 261 004 4 x 50 ml
High 2.5 AU Citrate and fluoride inhibit the enzyme activity.
261 005 2 x 50 ml
Sensitivity 5 U/L
Linearity 400 U/L Drugs
Reagent (R) Calcium dobesilate and doxycycline HCL cause artificially low
Tris buffer (pH 7.7 ) 80 mmol/L AST values at the tested drug level.
Procedure
L- Aspartate 240 mmol/L
MDH ≥ 450 U/L Macro Semi-Micro
LDH ≥ 1200 U/L Reagent (R) 1.0 ml 0.5 ml
Sodium Azide 8 mmol/L Specimen 100 ml 50 ml
NADH > 0.18 mmol/L
2 - Oxoglutarate 18 mmol/L Mix, read initial absorbance after 60 seconds. and start timer
Sodium Azide 8 mmol/L simultaneously. Read again after 60, 120 and 180 seconds.
Determine the mean absorbance change per minute (DA/min).
98 99
Alanine aminotransferase SYMBOLS IN PRODUCT LABELLING
Procedure Anticoagulants
Citrate and fluoride inhibit the enzyme activity.
Macro Semi-Micro
(ALT/GPT)-Liquizyme (4+1) EC REP Authorised Representative
Use by/Expiration Date Working 1.0 ml 500 ml Drugs
E.C.2.6.1.2. LOT
REF
Catalogue Number
for use
solution
Specimen 100 ml 50 ml
Calcium dobesilate and doxycycline HCL cause artificially low
ALT values at the tested drug level.
Mix, read initial absorbance after 60 seconds. and start timer Expected values
REF: 292 002 (10 x 10 ml) 100 test
REF: 292 003 ( 9 x 20 ml) 180 test Reagent 2 (R2 Coenzyme) simultaneously. Read again after 1, 2 and 3 minutes. Determine the o
mean absorbance change per minute (DA/min). 37 C Females up to 31 U/l (up to 0.52 mKat/L)
REF: 292 004 ( 4 x 60 ml) 240 test NADH ≥ 0.18 mmol/L males up to 41 U/l (up to 0.68 mKat/L)
REF: 292 005 ( 5 x 20 ml) 100 test 2 – Oxoglutarate 18 mmol/L
REF: 292 006 ( 4 x 50 ml) 200 test Sodium Azide 8 mmol/L Calculation o
REF: 291 007 ( 5 x100 ml) 500 test To calculate the ALT/GPT activity use the following formula
males up to 29 U/l (up to 0.48 mKat/L)
REF: 291 008 ( 6 x100 ml) 600 test For further information, refer to the Alanine aminotransferase
reagent material safety data sheet. U/l = 1780 x DA 334 nm /min o
Temperature conversion factor is 1.32 (25 30 C) and
Intended Use U/l = 1746 x DA 340 nm /min o
1.85 (25 37 C )
Spectrum Diagnostics liquizyme ALT reagent is intended for the in- Reagent preparation U/l = 3235 x DA 365 nm /min
vitro quantitative, diagnostic determination of ALT in human serum Prepare working solution as following:
on both automated and manual systems. Quality Control Spectrum Diagnostics does not interpret the results of a
Normal & abnormal control serum of known concentrations should clinical laboratory procedure; interpretation of the results is
Background be analyzed with each run. considered the responsibility of qualified medical personnel.
The enzyme alanine aminotransferase ALT is widely distributed with All indications of clinical significance are supported by
high concentrations in the liver and to a lesser extent in kidneys, Performance Characterstics literature references.
REF:292 004 : add12 ml from R2 to one bottle of R1; mix gently.
heart, skeletal muscles, pancreas and lungs. Elevated serum ALT REF:292 005 : add 4 ml from R2 to one bottle of R1; mix gently. Precision
is found in hepatitis, cirrhosis, obstructive jaundice, carcinoma of REF:292 006 : add10 ml from R2 to one bottle of R1; mix gently. Analytical Range
the liver, and chronic alcohol abuse. ALT is only slightly elevated in REF:292 007 : add one bottle of R2 to one bottle of R1;mix gently. Within run (Repeatiblity) 5 – 400 U/L.
patients who have an uncomplicated myocardial infarction. Although REF:292 008 : add one bottle of R2 to one bottle of R1; mix gently.
both serum aspartate aminotransferase AST and ALT become Level 1 Level 2
elevated whenever disease processes affect liver cell integrity,
Waste Disposal
Or prepare the working solution according to the number of tests n 20 20
ALT is the more liver specific enzyme. Moreover, elevations of ALT required by mixing 4 volumes of reagent 1 (R1) and 1volume of
Mean (U/L) 24.6 105.9 Please consult local regulations for a correct waste disposal.
activity persist longer than elevations of AST activity. reagent 2 (R2), e.g. 400 ml R1 + 100 ml R2.
SD 0.93 0.94
Method Precautions and Warnings CV% 3.78 0.89 S57: use appropriate container to avoid environmental
Kinetic method according to the International Federation of Clinical Do not ingest or inhalate. In case of contact with eyes or skin; rinse contamination.
Chemistry (IFCC) (3). immediately with plenty of soap and water. In case of severe injuries; S61: avoid release in environment. refer to special instructions/
seek medical advice immediately. Run to run (Reproducibility) safety data sheets.
Assay Principle
The series of the reaction involved in the assay system is as follows: Both reagents (R1) and (R2) contain sodium azide which may react Level 1 Level 2
References
with copper or lead plumbing. n 20 20
1. The amino group is enzymatically transferred by ALT present in 1. Breuer J, report on the symposium “drug effects in clinical
Mean (U/L) 25.2 106
the sample from alanine to the carbon atom of 2-oxoglutarate Reagent Storage and Stability chemistry methods”. Eur J Clin Chem Clin Biochem.
yielding pyruvate and L-glutamate. All reagents are stable until expiration date stated on label when SD 1.1 1.05 1996;34:385-386.
o
stored refrigerated at 2 - 8 C. Working solution is stable for 4 weeks CV% 3.9 0.95
L-Alanine Pyruvate o o 2. ECCLS. Determination of the catalytic activity concentration in
ALT at 2 – 8 C or 2 days at 15 - 25 C.
+ + serum on L- alanine aminotransferase (EC 2.6.1.2,ALAT) Clin
2-Oxoglutarate L-Glutamate chem. 1989;20:204-211.
Deterioration Methods Comparison
Do not use liquizyme ALT reagent if it is turbid or if the absorbance A comparison between Spectrum Diagnostics ALT (4+1) reagent 3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
2. Pyruvate is reduced to lactate by LDH present in the reagent of the working reagent is less than1.0 at 340 nm. Failure to recover and a commercial reagent of the same methodology was performed Biochem 1986;24:481-95 .
with the simultaneous oxidation of NADH to nicotinamide control values within the assigned range may be an indication of on 20 human sera. A correlation of 0.983 was obtained.
reagent deterioration. 4. Henry RJ, et al. Am J clin Path 1960 :34:381
adenine dinucleotide (NAD). The reaction is monitored by
measuring the rate of decrease in absorbance at 340 nm due Sensitivity 5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ, eds
to the oxidation of NADH. Specimen Collection and Preservation When run as recommended, the minimum detection limit of this Clinical chemistry, theory, analysis, and correlation. St
Use nonhemolyzed serum or plasma. Heparin and EDTA are the assay is 5.0 U/L. louis:mosby;1984:420-438.
Pyruvate L-Lactate only acceptable anticoagulants; avoid other anticoagulants. The
LDH + 6. Young DS. Effects of drugs on clinical laboratory tests. Third
+ biological half-life of ALT in serum is 47 hours.
NADH + H+ NAD+ o Linearity edition. 1990 :3:6-12.
Stability: 3 days at 15 - 25 C or 7 days
o o The reaction is linear up to ALT concentration of 400 U/L;
at either 4- 8 C or at -20 C 7. Zilva JF, pannall PR : plasma enzymes in diagnosis in
specimens showing higher concentration should be diluted 1+5 with
3. Endogenous sample pyruvate is rapidly and completely physiological saline and repeat the assay (result×6). clinical chemistry in diagnosis and treatment lioydluke london
reduced by LDH during the initial incubation period so that it System Parameters 1979:chap 17 : 338.
does not interfere with the assay. Wavelength 340 nm (334 – 365 nm)
Optical path 1 cm
Serum, plasma
Sample pyruvate L-Lactate Assay type Kinetic
LDH + ORDERING INFORMATION
+ Direction decrease
NAD+ Hemolysis
NADH + H+ Sample : Reagent Ratio 1 : 10 CATALOG NO QUANTITY
Erythrocyte contamination elevates results, since ALT activities
e.g .: Reagent volume 1 ml
in erythrocytes are 3 to 5 times higher than those in normal sera. 292 001 4 x 20 ml
o o 292 002 10 x 10 ml
Reagents Icterus 292 003 9 x 20 ml
Equilibration time 60 seconds.
No significant interference. 292 004 4 x 60 ml
Read time 1 to 3 minutes
Reagent 1 (R1 Buffer / Enzyme) 292 005 5 x 20 ml
Tris buffer (pH 7.4) 100 mmol/L Lipemia 292 006 4 x 50 ml
L- Alanine 800 mmol/L Lipemic specimens may cause high absorbance flagging. Diluted 292 007 5 x 100 ml
High 2.5 AU
LDH ≥ 2000 U/L sample is recommended. 292 008 6 x 100 ml
Sensitivity 5 U/L
Sodium Azide 8 mmol/L Linearity 400 U/L
100 101
Calculation
To calculate the ALT/GPT activity use the following formula
Expected values
(ALT/GPT)-Liquizyme (1 + 1)
o
EC REP Authorised Representative Temperature Limitation 37 C Females up to 31 U/l (up to 0.52 mKat/L)
IVD For in-vitro diagnostic use Use by/Expiration Date U/l = 1780 x DA 334 nm /min males up to 41 U/l (up to 0.68 mKat/L)
E.C.2.6.1.1. LOT
REF
Catalogue Number
for use
U/l = 1746 x DA 340 nm /min
U/l = 3235 x DA 365 nm /min 30 C
o
Females up to 22 U/l (up to 0.37 mKat/L)
males up to 29 U/l (up to 0.48 mKat/L)
REF: 263 001 (2 x 25 ml) 50 test
Quality Control
REF: 263 002 (4 x 25 ml) 100 test o
Normal & abnormal commercial control serum of known Temperature conversion factor is 1.32 (25 30 C ) and 1.85
REF: 263 003 (2 x 100 ml) 200 test o
concentrations should be analyzed with each run. (25 37 C )
Reagent 2 (R2 Coenzyme)
NADH ≥ 0.06 mmol/L
Performance Characterstics Spectrum Diagnostics does not interpret the results of a
Intended Use 2-Oxoglutarate 4 mmol/L
Spectrum Diagnostics liquizyme ALT reagent is intended for the in- Sodium Azide 8 mmol/L
Precision clinical laboratory procedure; interpretation of the results is
vitro quantitative, diagnostic determination of ALT in human serum considered the responsibility of qualified medical personnel.
on both automated and manual systems. For further information, refer to the Alanine aminotransferase Within run (Repeatiblity) All indications of clinical significance are supported by
reagent material safety data sheet. literature references.
Level 1 Level 2
Background
n 20 20
The enzyme alanine aminotransferase ALT is widely distributed with Precautions and Warnings Analytical Range
high concentrations in the liver and to a lesser extent in kidn- e y s , Do not ingest or inhalate. In case of contact with eyes or skin; rinse Mean (U/L) 103 190 5 – 400 U/L.
heart, skeletal muscles, pancreas and lungs. Elevated serum ALT immediately with plenty of soap and water. In case of severe injuries; SD 6.1 13
is found in hepatitis, cirrhosis, obstructive jaundice, carcinoma of seek medical advice immediately.
CV% 6 7.4 Waste Disposal
the liver, and chronic alcohol abuse. ALT is only slightly elevated in
patients who have an uncomplicated myocardial infarction. Both reagents (R1) and (R2) contain sodium azide which may react
Although both serum aspartate aminotransferase AST and ALT with copper or lead plumbing.
Run to run (Reproducibility) S56: dispose of this material and its container at hazardous or
become elevated whenever disease processes affect liver cell
integrity, ALT is the more liver specific enzyme. Moreover, eleva- Reagent Preparation, Storage and Stability Level 1 Level 2 S57: use appropriate container to avoid environmental
tions of ALT activity persist longer than elevations of AST activity. All reagents are stable until expiration date stated on label when
o n 20 20 contamination.
stored refrigerated at 2 - 8 C. S61: avoid release in environment. refer to special instructions/
Method Working solution can be prepared by adding equal volumes Mean (U/L) 103 190 safety data sheets.
o
Kinetic method according to the International Federation of from R1 and R2 , Stability: 2 days 2 – 8 C.
SD 14.8 16
ClinicalChemistry (IFCC) (3).
CV% 14.3 8
References
Deterioration 1. Breuer J, report on the symposium “drug effects in clinical
Assay Principle Do not use liquizyme ALT reagent if it is turbid or if the absorbanceof chemistry methods”. Eur J Clin Chem Clin. 1996;34:385-386.
The series of the reaction involved in the assay system is as follows: the working reagent is less than 1.0 at 340 nm.
Failure to recover control values within the assigned range Methods Comparison 2. ECCLS. Determination of the catalytic activity concentration in
1. The amino group is enzymatically transferred by ALT present in may be an indication of reagent deterioration. A comparison between Spectrum Diagnostics ALT (1+1) reagent serum on L- alanine aminotransferase (EC 2.6.1.2,ALAT) Clin
the sample from alanine to the carbon atom of 2-oxoglutarate and a commercial reagent of the same methodology was performed chem. 1989;20:204-211.
yielding pyruvate and L-glutamate. Specimen Collection and Handling on 20 human sera.A correlation of 0.997 was obtained.
3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
Use nonhaemolyzed serum or plasma. Heparin and EDTA are the Biochem 1986;24:481-95.
L-Alanine Pyruvate Sensitivity
ALT + only acceptable anticoagulants; avoid other anticoagulants. The
+ biological half-life of ALT in serum is 47 hours. When run as recommended, the minimum detection limit of this 4. Henry RJ, et al. Am J clin Path 1960 :34:381.
2-Oxoglutarate L-Glutamate
assay is 5.0 U/L.
o 5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ, eds.
Stability: 3 days at 15 - 25 C or 7 days at
Clinical chemistry, theory,analysis, and correlation. St
o
either 4- 8oC or at - 20 C Linearity
louis:mosby;1984:420- 438.
2. Pyruvate is reduced to lactate by LDH present in the reagent The reaction is linear up to ALT concentration of 400 U/L;
with the simultaneous oxidation of NADH to nicotinamide System Parameters specimens showing higher concentration should be diluted 1+5 with 6. Young DS. Effects of drugs on clinical laboratory tests. Third
adenine dinucleotide (NAD). The reaction is monitored by Wavelength 340 nm (334 – 365 nm) physiological saline and repeat the assay (result×6). edition. 1990 :3:6-12.
measuring the rate of decrease in absorbance at 340 nm due Optical path 1 cm
to the oxidation of NADH. Assay type Kinetic Interfering Substances 7. Zilva JF, pannall PR : plasma enzymes in diagnosis in clinical
Direction decrease Serum, plasma chemistry in diagnosis and treatment lioyd-luke london
Pyruvate L-Lactate Sample: Reagent Ratio 1 : 10 1979:chap 17 : 338.
LDH +
+ e.g .: Reagent volume 1 ml Haemolysis
NADH + H+ NAD+ Sample volume 100 ml Erythrocyte contamination elevates results, since ALT activities
o o
in erythrocytes are 3 to 5 times higher than those in normal sera. ORDERING INFORMATION
Equilibration time 30 seconds. CATALOG NO QUANTITY
3. Endogenous sample pyruvate is rapidly and completely Read time 1 to 3 minutes Icterus
263 001 2 x 25 ml
reduced by LDH during the initial incubation period so that it Zero adjustment Against air No significant interference.
263 002 4 x 25 ml
does not interfere with the assay. Reagent Blank Limits Low 1.00 AU
263 003 2 x 100 ml
High 2.5 AU Lipemia
Sample pyruvate L-Lactate Sensitivity 5 U/L Lipemic specimens may cause high absorbance flagging. Diluted
LDH +
+ Linearity 400 U/L sample is recommended.
NADH + H+ NAD+
Procedure Anticoagulants
Citrate and fluoride inhibit the enzyme activity.
Pipette in a test tube:
Reagents Working solution 1 ml (Or 0.5 ml R1 + 0.5 ml R2) Drugs
Specimen 100 ml Calcium dobesilate and doxycycline HCL cause artificially low
Reagent 1 (R1 Buffer / Enzyme) ALT values at the tested drug level.
Tris buffer ( pH 7.4) 100 mmol/L Mix, read initial absorbance after 30 seconds. and start timer
L- Alanine 1.4 mol/L simultaneously. Read again after 1, 2 and 3 minutes. Determine
LDH ≥ 3500 U/L the mean absorbance change per minute (DA/min).
Sodium Azide 0.06 mmol/L
102 103
Calculation
The ALT activity in the serum can be determined from the following
References
1. Henry RJ et al. Am J Clin Path 1960 :34:381.
(ALT/GPT) - Colorimetric EC REP Authorised Representative
table:
2. Reitman S and Frankel S . Am . J.Clin.Path., 1975 ;28;65.
LOT Batch Code/Lot number for use Absorbance U/L Absorbance U/L 3. Sherwin JE. Liver function. In:kaplan LA, Pesce AJ, eds.
REF: 264 001 ( 2 x 50 ml ) 100 test REF Catalogue Number Manufactured by
0.025 4 0.275 48 Clinical chemistry, theory, analysis, and correlation. St
REF: 264 002 ( 2 x 100 ml ) 200 test Consult instructions for use (C) - Corrosive
0.050 8 0.300 52 louis:Mosby;1984:420-438.
0.075 12 0.325 57
4. Young DS. Effects of drugs on clinical laboratory tests. Third
0.100 17 0.350 62
Intended Use edition. 1990 :3:6-12.
Reagent Preparation, Storage and Stability 0.125 21 0.375 67
Spectrum Diagnostics ALT reagent is intended for the in-vitro The reagents are supplied ready-to-use and stable up to the expiry 0.150 25 0.400 72
quantitative, diagnostic determination of ALT in human serum. o
date labeled on the bottles when stored at 2 – 8 C. 0.175 29 0.425 77
0.200 34 0.450 83 ORDERING INFORMATION
Background Deterioration 0.225 39 0.475 88 CATALOG NO QUANTITY
The enzyme alanine aminotransferase (ALT) is widely distributedwith Do not use the ALT regents if precipitate forms. Failure to recover 0.250 43 0.500 94
high concentrations in liver and to a lesser extent in kidneys, heart, 264 001 2 x 50 ml
skeletal muscles, pancreas and lungs. Elevated serum ALT is found Quality Control 264 002 2 x 100 ml
in hepatitis, cirrhosis, obstructive jaundice, carcinoma of the liver, Normal & abnormal commercial control serum of known
and chronic alcohol abuse. ALT is only slightly elevated in patients Specimen Collection and Preservation concentrations should be analyzed with each run.
who have an uncomplicated myocardial infarction. Although both Use only non haemolyzed serum. The biological half-life of ALT in
serum aspartate aminotransferase (AST) and ALT become elevated serum is 47 hours. The only acceptable anticoagulants are heparin Sensitivity
whenever disease processes affect liver cell integrity, ALT is the and EDTA When run as recommended, the sensitivity of this assay is 4 U/L.
more liver specific enzyme. Moreover,elevations of ALT activity o
Stability: 3 days at 15 - 25 C or 7 days
persist longer than elevations of AST activity. o o
at either 4 - 8 C or at -20 C Linearity
The assay is linear up to 94 U/L. If the absorbance exceeds 0.5 at
Method System Parameters 546 nm (ALT 94 U/L) samples should be diluted 1 + 9 using sodium
ALT – (colorimetric method). Wavelength 546 nm (530-550 nm) chloride and repeat the assay (result × 10).
Optical path 1 cm
Assay Principle Assay type End-point Interfering Substances
The reaction involved in the assay system is as follows: Direction Increase Serum, plasma
The amino group is enzymatically transferred by ALT present in the Sample: Reagent Ratio 1 : 60
sample from alanine to the carbon atom of 2-oxoglutarate yielding o
Temperature 37 C and 20 – 25 C
o
Haemolysis
pyruvate and L-glutamate. Zero adjustment Reagent or Sample blank Erythrocyte contamination may elevate results, since ALT activities
Sensitivity 4 U/L in erythrocytes are three to five times than those in normal sera.
L-Alanine Pyruvate
ALT + Linearity 94 U/L
+
2-Oxoglutarate L-Glutamate Icterus
Procedure No significant interference.
ALT activity is measured by monitoring the concentration of pyruvate 1. Measurement against Reagent Blank Lipemia
hydrazone formed with 2,4-dinitrophenylhydrazine. Lipemic specimens may cause high absorbance flagging. Diluted
Pipette into test tubes sample is recommended.
Reagents Reagent blank Sample
Note
Reagent 1 (R1 Buffer) R1(buffer) 0.5 ml 0.5 ml High concentration of aldehydes, ketones, or oxo-acids in some
Phosphate buffer 100 mmol/L Sample ------ 100 ml sera may cause false high transaminase levels. Measurement
DL- Alanine 200 mmol/L Distilled water 100 ml ------ aganist a serum blank instead of a reagent blank avoids the risk of
2 – Oxoglutarate 6 mmol/L o finding such artifacts.
Mix and incubate for exactly 30 minutes at 37 C
Sodium Azide 12 mmol/L
R2 0.5 ml 0.5 ml Expected Values
Reagent 2 (R2) Mix and incubate for exactly 20 minutes at 20 – 25 C
o Serum : up to 12 U/L.
2,4-dinitrophenylhydrazine 2.0 mmo/L
(C)-Corrosive contains caustic materials. Sodium hydroxide 5.0 ml 5.0 ml
R35 Causes severe burns. Mix, measure absorbance of specimen against reagent blank at clinical laboratory procedure; interpretation of the results is
R41 Risk of serious damage to eyes. 546 nm after 5 minutes. considered the responsibility of qualified medical personnel.
S26 In case of contact with eyes, rinse immediately All indications of clinical significance are supported by
with plenty of water and seek medical advice. 2. Measurement against Sample Blank literature references.
S28 After contact with skin, wash immediately with
plenty of soap and water. Sample blank Sample
For further information, refer to the Alanine aminotransferase R1(buffer) 0.5 ml 0.5 ml Analytical Range
reagent material safety data sheet. Sample ----- 100 ml 4 – 94 U/L.
o
Mix and incubate for exactly 30 minutes at 37 C
Precautions and Warnings Waste Disposal
Do not ingest or inhalate. In case of contact with eyes or skin; rinse R2 0.5 ml 0.5 ml
immediately with plenty of soap and water. In case of severe injuries; Sample 100 ml ----- Please consult local regulations for a correct waste disposal.
seek medical advice immediately. Mix and incubate for exactly 20 minutes at 20 – 25 C
o
Reagent (R1) contains sodium azide which may react with copper Sodium hydroxide 5.0 ml 5.0 ml
or lead plumbing. Mix, measure absorbance of specimen against sample blank at contamination.
546 nm after 5 minutes. S61: avoid release in environment. refer to special instructions/
Additional Reagent safety data sheets.
Sodium hydroxide 0.4 mol/L.
104 105
Quality Control
(ALT/GPT) - Ultimate Single Reagent EC REP Authorised Representative Temperature Limitation be analyzed with each run.
E.C.2.6.1.2. LOT Batch Code/Lot number CAUTION. Consult instructions Performance Characterstics literature references.
REF Catalogue Number for use Precision
REF: 265 002 (6 x 20 ml) 120 test Within run (Repeatiblity)
Analytical Range
REF: 265 003 (2 x 100 ml) 200 test 5 – 400 U/L.
REF: 265 004 (4 x 50 ml) 200 test Level 1 Level 2
For further information, refer to the Alanine aminotransferase Waste Disposal
REF: 265 005 (2 x 50 ml) 100 test n 20 20
reagent material safety data sheet. This product is made to be used in professional laboratories.
Mean (U/L) 24.6 105.9 Please consult local regulations for a correct waste disposal.
Intended Use
Spectrum Diagnostics Ultimate ALT reagent is intended for the in-
Reagent Preparation, Storage and Stability SD 0.93 0.94 S56: dispose of this material and its container at hazardous or
Spectrum Ultimate ALT reagent is supplied ready-to-use and stable special waste collection point.
vitro quantitative, diagnostic determination of ALT in human serum CV% 3.78 0.89
up to the expiry date labeled on the bottles. S57: use appropriate container to avoid environmental
on both automated and manual systems.
Once opened, the opened vial is stable for 3 months at the specified contamination.
temperature. S61: avoid release in environment. refer to special instructions/
Background Run to run (Reproducibility)
The enzyme alanine aminotransferase ALT is widely distributed with safety data sheets.
high concentrations in the liver and to a lesser extent in kidneys,
Precautions and Warnings Level 1 Level 2
Do not ingest or inhalate. In case of contact with eyes or skin; rinse References
heart, skeletal muscles, pancreas and lungs. Elevated serum ALT n 20 20
is found in hepatitis, cirrhosis, obstructive jaundice, carcinoma of
seek medical advice immediately. Mean (U/L) 25.2 106
the liver, and chronic alcohol abuse. ALT is only slightly elevated in 1. Breuer J, report on the symposium “drug effects in clinical
patients who have an uncomplicated myocardial infarction. Although SD 1.1 1.05 chemistry methods”. Eur J Clin Chem Clin Biochem.
The reagent (R) contain sodium azide which may react with copper
both serum aspartate aminotransferase AST and ALT become CV% 3.9 0.95 1996;34:385-386.
or lead plumbing.
elevated whenever disease processes affect liver cell integrity,
ALT is the more liver specific enzyme. Moreover, elevations of ALT 2. ECCLS. Determination of the catalytic activity concentration in
activity persist longer than elevations of AST activity. Deterioration Methods Comparison serum on L- alanine aminotransferase (EC 2.6.1.2,ALAT) Clin
Do not use Spectrum Ultimate ALT reagent if it is turbid or if the A comparison between Spectrum Diagnostics ALT reagent and a chem. 1989;20:204-211.
absorbance of the working reagent is less than 0.9 at 340 nm. commercial reagent of the same methodology was performed on 20
Method human sera. A correlation of 0.983 was obtained. 3. IFCC expert panel on enzymes part 3. J Clin Chem Clin
Failure to recover control values within the assigned range may be
Kinetic method according to the International Federation of Clinical Biochem 1986;24:481-95 .
an indication of reagent deterioration.
Chemistry (IFCC) (3).
Sensitivity 4. Henry RJ, et al. Am J clin Path 1960 :34:381
Specimen Collection and Preservation When run as recommended, the minimum detection limit of this
Assay Principle assay is 5.0 U/L.
Use nonhemolyzed serum or plasma. Heparin and EDTA are the 5. Sherwin JE. Liver function. In:kaplan LA, PESCE AJ, eds.
The series of the reaction involved in the assay system is as follows:
only acceptable anticoagulants; avoid other anticoagulants. The Clinical chemistry, theory, analysis, and correlation. St
1. The amino group is enzymatically transferred by ALT present in
biological half-life of ALT in serum is 47 hours. Linearity louis:mosby;1984:420-438.
o
Stability: 3 days at 15 - 25 C or 7 days The reaction is linear up to ALT concentration of 400 U/L;
the sample from alanine to the carbon atom of 2-oxoglutarate o o 6. Young DS. Effects of drugs on clinical laboratory tests. Third
at either 4- 8 C or at -20 C specimens showing higher concentration should be diluted 1+5 with
yielding pyruvate and L-glutamate. edition. 1990 :3:6-12.
physiological saline and repeat the assay (result×6).
L-Alanine Pyruvate System Parameters 7. Zilva JF, pannall PR : plasma enzymes in diagnosis in
ALT
+ + Wavelength 340 nm (334 – 365 nm) Interfering Substances clinical chemistry in diagnosis and treatment lioydluke london
2-Oxoglutarate L-Glutamate Optical path 1 cm Serum, plasma 1979:chap 17 : 338.
Assay type Kinetic
Direction decrease Hemolysis
2. Pyruvate is reduced to lactate by LDH present in the reagent Sample : Reagent Ratio 1 : 10 Erythrocyte contamination elevates results, since ALT activities
with the simultaneous oxidation of NADH to nicotinamide e.g .: Reagent volume 1 ml in erythrocytes are 3 to 5 times higher than those in normal sera. ORDERING INFORMATION
adenine dinucleotide (NAD). The reaction is monitored by Sample volume 100 ml CATALOG NO QUANTITY
measuring the rate of decrease in absorbance at 340 nm due Temperature 37 C or 30 C
o o
Icterus
to the oxidation of NADH. 265 001 2 x 20 ml
Equilibration time 60 seconds No significant interference.
265 002 6 x 20 ml
L-Lactate Read time 180 seconds
Pyruvate LDH 265 003 2 x 100 ml
+ Zero adjustment Against air Lipemia
+ 265 004 4 x 50 ml
NAD+ Reagent Blank Limits Low 0.9 AU Lipemic specimens may cause high absorbance flagging. Diluted
NADH + H+ 265 005 2 x 50 ml
High 2.5 AU sample is recommended.
Sensitivity 5 U/L
3. Endogenous sample pyruvate is rapidly and completely Linearity 400 U/L Anticoagulants
reduced by LDH during the initial incubation period so that it Citrate and fluoride inhibit the enzyme activity.
does not interfere with the assay. Procedure:
Drugs
Sample pyruvate L-Lactate Macro Semi-Micro Calcium dobesilate and doxycycline HCL cause artificially low
LDH +
+ Reagent (R) 1.0 ml 500 ml ALT values at the tested drug level.
NADH + H+ NAD+
Specimen 100 ml 50 ml
Expected values
Mix, read initial absorbance after 60 seconds. and start timer o
Reagent (R) simultaneously. Read again after 60, 120 and 180 seconds. 37 C Females up to 31 U/l (up to 0.52 mKat/L)
Tris buffer (pH 7.4) 100 mmol/L Determine the mean absorbance change per minute (DA/min). males up to 41 U/l (up to 0.68 mKat/L)
L- Alanine 800 mmol/L o
LDH ≥ 2000 U/L 30 C Females up to 22 U/l (up to 0.37 mKat/L)
Calculation
Sodium Azide 8 mmol/L males up to 29 U/l (up to 0.48 mKat/L)
To calculate the ALT/GPT activity use the following formula
NADH ≥ 0.18 mmol/L o
2 – Oxoglutarate 18 mmol/L Temperature conversion factor is 1.32 (25 30 C) and
U/l = 1780 x DA 334 nm /min o
Sodium Azide 8 mmol/L 1.85 (25 37 C )
U/l = 1746 x DA 340 nm /min
U/l = 3235 x DA 365 nm /min
The reagent also contains additives required to maintain NADH in
its reduced forrm.
106 107
Glucose-6-Phosphate SYMBOLS IN PRODUCT LABELLING
2. Place cuvette in constant temperature compartment or water
bath and incubate for approximately 5 minutes to obtain
Linearity
The assay is linear up to 19.5 m/g Hb
dehydrogenase (G-6-PDH) EC REP Authorised Representative
thermal equilibrium.
Expected Values
3. Read and record absorbance (A) of Test at 340nm vs water or
REF: 253 001 10 tests LOT Batch Code/Lot number CAUTION. Consult instructions Pottasium Dichromate solution. This is INITIAL A. (if using the G6PDH Activity (U/g Hb.): 4.6 -13.5 at 30°C
REF Catalogue Number for use water bath or incubator, return the cuvet to it) 6.4 -18.7 at 37°C
Reagent 1: Assay Reagent 10 ml Consult instructions for use Manufactured by (U/10¹² RBC’s): 146 - 376 at 30°C
4. Exactly 5 minutes later, again read and record absorbance. 202 - 522 at 37°C
Reagent 2 : Substrate Reagent 20 ml
This is FINAL A. Note:
The integrity of erythrocytes collected in ACD is preserved even
It is recommended for each laboratory to establish and maintain its
after prolonged storage so that obtaining accurate red cell counts 5. To determine G-6-PDH activity, refer to “calculations” section.
Intended Use own reference values. The given data are only an indication.
usually poses no problem. However, red cell counts on specimens
Spectrum-Diagnostics G-6-PDH reagent is intended for the in-vitro collected in heparin become unreliable after about 2 days. Thus, CALCULATION
quantitative UV diagnostic estimation of G-6-PDH in human blood. for heparinized samples, results are best reported in terms of Spectrum Diagnostics does not interpret the results of a
hemoglobin concentration. Both copper, which completely inhibits FINAL A - INITIAL clinical laboratory procedure; interpretation of the results is
A per min =
Background the enzyme at a concentration of 100ìmol/L, and sulfate ions (0.005 5 considered the responsibility of qualified medical personnel.
Glucose-6-Phosphate-Dehydrogenase (G6PDH) deficiency is one mol/L) will decrease observed values of G-6-PDH activity, certain G-6-PDH activity is expressed as U/1012 erythrocytes (RBC) or U/g All indications of clinical significance are supported by
of the most common human enzyme deficiencies in the world. drugs and other substances are known to influence circulating hemoglobin (Hb). literature references.
During G6PD deficiency, the red cells are unable to regenerate levels of G-6-PDH. Reticulocytes have higher G-6-PDH levels than
reduced Nicotineamide adenine dinucleotide phosphate (NADPH), mature red cells. Therefore it is not recommended that assays be 48,390
G-6-PDH (U/1012 RBC) = A per min x x TCF
a reaction that is normally catalyzed by the G6PD enzyme. Since the performed after a severe hemolytic crisis, since G-6-PDH levels N Waste Disposal
X chromosome carries the gene for G6PD enzyme, this deficiency appear falsely elevated. Under those conditions, detection of This product is made to be used in professional laboratories.
Where :
mostly affects the males. The two major conditions associated with deficiency may require family studies. Testing may be more helpful Please consult local regulations for a correct waste disposal.
G6PD deficiency are hemolytic anaemias and neonatal jaundice, after the level of mature red cells has returned to normal. Under S56: dispose of this material and its container at hazardous or
N = Red cell count divided by 106
which may result in neurological complications and death. Screening normal circumstances, activity contributed by leucocytes, platelets special waste collection point.
TCF = Temperature correction factor (1 at 30 °C)
and detection of G6PD deficiency helps in reducing such episodes, and serum is relatively small. However, in cases of extreme anemia, S57: use appropriate container to avoid environmental
through appropriate selection of treatment, patient counseling and grossly elevated white counts or very low levels of red cell G-6-PDH 48390 x TCF contamination.
G-6-PDH (U/g Hb) = A per min x
abstinence from disease precipitating drugs such as anti malarials activity, the contribution to the total made under these conditions Hb(g/dL) S61: avoid release in environment. refer to special instructions/
and other agents. may be significant. See “Use of Buffy-Coat-Free Samples” section. safety data sheets.
EXAMPLE :
Method Reagent preparation Assay of a specimen which had a red cell count of 4.6 x /mm3 References
UV-Kinetic Method. G-6-PDH Reagents are supplied ready to use. and a hemoglobin concentration 15.2 g/dL resulted in a A per min S.K. Sood et al., The Indian journal of path and micro,, 24 (1981), 89.
at 30°C of 0.026. Lubin, B.H. and Oski, F.A., J. Pediatr. 70 (1967), 788.
Assay Principle System Parameters
G6PDH in the RBC’s is released by a lysing agent present in the Wavelength 340 nm G-6-PDH(U/1012 RBC) = 0.026 x 48,390 = 295
reagent. The G6PDH released catalyzes the oxidation of Glucose Optical path 1 cm 4.6
Assay type UV-Kinetic 4839 CATALOG NO QUANTITY
6 phosphate with the reduction of NADP to NADPH. The rate G-6-PDH(U/g Hb) = 0.026 x = 8.9
Direction Increase 15.2 253 001 10 Test
of reduction of NADP to NADPH is measured as an increase
in absorbance which is proportional to the G6PDH activity in the Sample: Reagent Ratio 1:100 Note: If A per min is greater than 0.060, repeat determination (3)
o
sample. Temperature 30 C using 5ìL blood and multiply results by 2
Measurement Against distilled water
G-6-PDH Delay/Lag/Time 300 sec CALIBRATION
G-6-P +NADP Gluconate -6-P+ NADPH+H Interval Time 60 sec The procedure is standardized on the basis of the milimolar
NO. OF READINGS 05 absorptivity of NADPH, which is 6.22 at 340nm. The oxidative
Reagents Blank Absobance Limit < 0.8 conversion of G-6-P by G-6-PDH leads to reduction of NADP to
Reagent 1 (R1) : Assay Reagent Factor 4839 NADPH on a molar equivalent basis. Measurement of the rate of
Reagent 2 (R2) : Substrate Reagent Low Normal at 30°C 4.6 m/g Hb increase in absorbance ( A) at340nm serves to quantitate enzymatic
G6PDH Control (Vial) High Normal at 30°C 13.5 m/g Hb activity. The maximum G-6-PDH activity which may be measured by
Linearity at 30°C 19.5 m/g Hb this procedure is approximately 650 U/1012 RBC or 19.5 U/g Hb.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Procedure USE OF BUFFY-COAT-FREE SAMPLE
immediately with plenty of soap and water. In case of severe injuries; The temperature of the reaction mixture should be maintained Under normal circumstances G-6-PDH activity contributed by
seek medical advice immediately. at 30°C or some other constant temperature (see “Temperature leucocytes, platelets and serum is relatively small. However, as
Correction” section). reported by Echler and others, more accurate measurement of
Reagent Storage and Stability G-6-PDH activity, specially in the presence of anaemia and /
o
Reagents and standard are ready-to-use. When stored at 2 – 8 C; 1. Prepare reaction mixture : or leucocytosis, can be achieved by using buffy coat-free blood
they are stable up to the expiry date stated on the label. samples for assay. Thus in case of a boderline value obtained with
Recontituted G6PDH Assay solution is stable for 8hrs at room temp. b) Add 0.01ml blood to 1.0 ml of G-6-PDH Assay reagent and mix
thoroughly to completely suspend erythrocytes. Let stand at whole blood, it may be warranted to repeat the assay on a buffy
(15 - 25 °C) or 5 days refrigerated (2 - 8 °C). coat-free sample.
room temperature (18-26°C) for 5-10 minutes.
Sample collection and preparation c) Add 2.0ml G-6-PDH Substrate Reagent and mix gently by TEMPERATURE CORRECTION
Whole blood collected in EDTA, Heparin or ACD is satisfactory. Red inverting several times. When temperature of 30°C, no temperature correction factor (TCF)
cell G-6-PDH is stable in whole blood for one week refrigerated (2- is required in the calculations. If assay is performed at a room
8°C), but is unstable in red cell hemolysates. Freezing of blood is d) Transfer contents to cuvette labeled Test & proceed with Step2.
temperature other than 30°C, a TCF must be used. When the
not recommended. Since activity is reported in terms of number of temperature is 37° C, the TCF is 0.66.
red cells or grams of hemoglobin. The red cell count or hemoglobin
concentration should be determined prior to performing the G-6-
PDH assay.
108 109
Glucose-6-Phosphate SYMBOLS IN PRODUCT LABELLING
2.G6PDH Activity (U/g Hb)

224 x 100 clinical laboratory procedure; interpretation of the results is
dehydrogenase plus (G-6-PDH plus) EC REP Authorised Representative Temperature Limitation = DA /Min X
6.22 x Hb (g/dl) considered the responsibility of qualified medical personnel.
REF: 372 001 (10 x1.1 ml) 10 tests LOT Batch Code/Lot number CAUTION. Consult instructions
Reagent 1: G6PDH (Co-enzyme-substrate) 10 vials REF Catalogue Number for use 3601
Reagent 2: G6PDH (Buffer) 12 ml = DA /Min X
Consult instructions for use Manufactured by Hb (g/dl)
Reagent 3 : G6PDH (Lysis reagent) 5.5 ml Waste Disposal
Where 100 = Factor to convert to 100 ml This product is made to be used in professional laboratories.
REF: 372 002 (20 x1.1 ml) 20 tests 224 = Total assay volume to sample volume Please consult local regulations for a correct waste disposal.
Reagent 1: G6PDH (Co-enzyme-substrate) 20 vials
Preparation of Red cell hemolysate 6.22 = Millimolar absorptivity of NADPH at 340 nm S56: dispose of this material and its container at hazardous or
Wash 0.1 ml of whole blood with 2 ml aliquots of physiological saline Hb (g/dl) = Hemoglobin concentration
Reagent 2: G6PDH (Buffer) 24 ml special waste collection point.
(0.9 %) 3 times and suspend the washed, packed and centrifuged
Reagent 3 : G6PDH (Lysis reagent) 11 ml S57: use appropriate container to avoid environmental
erythrocytes in precooled 0.5 ml of G-6-PDH Lysing reagent (R3), EXAMPLE :
contamination.
Mix well and keep in the refrigerator (2 - 4 °C) for at least 15 minutes Assay of a specimen which had a red cell count of 4.6 x106 /mm3 S61: avoid release in environment. refer to special instructions/
and maximum for 2 hours. and a hemoglobin concentration 15.2 g/dL resulted in a DA /Min
Intended Use Centrifuge the lysate at 3000 r.p.m, for 5 minutes prior to use.
safety data sheets.
Spectrum-Diagnostics G-6-PDH reagent is intended for the in-vitro at 30°C of 0.028.
quantitative estimation of G-6-PDH in erythrocytes. 36013 References
preparation of Reagent G-6-PDH(U/10¹² RBC) = 0.028 x = 219
Add 1.1 ml G-6-PDH buffer (R2) to the vial labelled G-6-PDH (R1). 4.6 1. Gross R.T. Hurwitz R.E.Marks P.A. Red cell Glucose-6-
Background Mix well and use after 5 minutes. 3601 phosphate dehydrogenase deficiency, J.Clin.Invest (1978)
G-6-PDH(U/g Hb) = 0.028 x = 6.83
Glucose-6-Phosphate-Dehydrogenase (G6PDH) deficiency is one 15.2 37.176.
of the most common human enzyme deficiencies in the world. 2. Kachmar J.F Moss.D.W,Enzymes,In Fundamentals of clinical
During G6PD deficiency, the red cells are unable to regenerate
System Parameters CALIBRATION chemistry Ed, by N.W. Teitz,Saunders philadelphia 1976 pp
Wavelength 340 nm The procedure is standardized on the basis of the milimolar
reduced Nicotineamide adenine dinucleotide phosphate (NADPH), 666-672.
Optical path 1 cm absorptivity of NADPH, which is 6.22 at 340nm. The oxidative
a reaction that is normally catalyzed by the G6PD enzyme. Since
Assay type Kinetic/ Increasing OD conversion of G-6-P by G-6-PDH leads to reduction of NADP to
the X chromosome carries the gene for G6PD enzyme, this
Direction Increase NADPH on a molar equivalent basis. Measurement of the rate of
deficiency mostly affects the males. The two major conditions
Temperature 30 °C increase in absorbance ( A) at 340nm serves to quantitate enzymatic ORDERING INFORMATION
associated with G6PD deficiency are hemolytic anaemias and
Measurement Against distilled water activity. The maximum G-6-PDH activity which may be measured by CATALOG NO QUANTITY
neonatal jaundice, which may result in neurological complications
Delay/Lag/Time 90 sec this procedure is approximately 650 U/10¹² RBC or 19.5 U/g Hb.
and death. Screening and detection of G6PD deficiency helps in 372 001 10 x 1.1ml
Interval Time 60 sec
reducing such episodes, through appropriate selection of treatment, 372 002 20 x 1.1ml
NO. of reading 4 USE OF BUFFY-COAT-FREE SAMPLE
patient counseling and abstinence from disease precipitating drugs
sample volume 25 ml (0.025 ml) Under normal circumstances G-6-PDH activity contributed by
such as anti malarials and other agents.
working reagent volume 1.0 ml leucocytes, platelets and serum is relatively small. However, as
Low Normal at 30°C 4.6 m/g Hb reported by Echler and others, more accurate measurement of
Method High Normal at 30°C 13.5 m/g Hb
UV-Kinetic Method. G-6-PDH activity, specially in the presence of anaemia and /
Linearity at 30°C 19.5 m/g Hb or leucocytosis, can be achieved by using buffy coat-free blood
samples for assay. Thus in case of a borderline value obtained with
Assay Principle Procedure whole blood, it may be warranted to repeat the assay on a buffy
G6PDH in the RBC’s is released by a lysing agent present in the
Pipette into test tubes coat-free sample.
reagent. The G6PDH released catalyzes the oxidation of Glucose
6 phosphate with the reduction of NADP to NADPH. The rate Working solution 1.0 ml
of reduction of NADP to NADPH is measured as an increase in TEMPERATURE CORRECTION
absorbance which is proportional to the G6PDH activity in the Hemolysate 0.025 ml When temperature of 30°C, no temperature correction factor (TCF)
sample. is required in the calculations. If assay is performed at a room
Add 1ml of working solution to 0.025 ml of hemolysate , mix and temperature other than 30°C, a TCF must be used. When the
G-6-PDH aspirate. After the intial delay of 90 seconds,record the absorbance temperature is 37°C, the TCF is 0.66.
G-6-P +NADP Gluconate -6-P+ NADPH+H of test at the interval of 1 minute for the next 3 minute at 340
nm. Determine the mean change in absorbance per minute and Linearity
Reagents calculate test results. The assay is linear up to 19.5 m/g Hb
Reagent 1 (R1): G6PDH ( Co-enzyme-substrate)
Reagent 2 (R2): G6PDH (Buffer) CALCULATION Expected Values
Reagent 3 (R3) : G6PDH (Lysis reagent)
G6PDH Activity (U/g Hb.): 4.6 - 13.5 at 30°C
1. G6PDH Activity (U/10¹² RBC’s): 6.4 - 18.7 at 37°C
Precautions and Warnings (U/10¹² RBC’s): 146 - 376 at 30°C
The Lysing Reagent (G6PDH) (R3) should be used cold (0 - 8 °C) 224 x 1012
= DA /Min X 202 - 522 at 37°C
to avoide the decrease in enzyme activity. 6.22 x N x 106 x 1000
Note:
36013 It is recommended for each laboratory to establish and maintain its
Reagent Storage and Stability = DA /Min X own reference values. The given data are only an indication.
N
G6PDH reagents are stable until the expiry date stated on the label. If the G6PDH activity is very low , measure the absorbance change
Reconstituted reagent is stable for 1 week at (2 - 8 °C). Where 224 = Total assay volume to sample volume for 5 minute after addition of buffered substrate and divide by 5 to
1012 = Factor for expressing activity in 1012 cells obtain DA /minute and calculate the test results.
Sample collection and preparation 6.22 = Millimolar absorptivity of NADPH at 340 nm
Fresh whole blood is the specimen required.Collection of blood Nx106 = Number of erythrocytes/cmm
by using any one of the anticoagulants such as citrate,oxalate or 1000 = Conversion of cell count from count per cmm
heparin is recommended. to count per ml.
Determine the Hemoglobin content of the whole blood and the
RBC’s count prior to lysis of the cells.
110 111
Lactate dehydrogenase SYMBOLS IN PRODUCT LABELLING
Precision
Analytical Range:
10 - 1200 U/L.
(LDH)-Liquizyme (4+1) EC REP Authorised Representative
Use by/Expiration Date Within run (Repeatiblity) Waste Disposal
E.C.1.1.1.27. LOT
REF
Catalogue Number
for use
Level 1 Level 2
REF: 283 001 ( 4 x 20 ml) 80 test Consult instructions for use Manufactured by n 20 20 S56: dispose of this material and its container at hazardous or
REF: 283 002 (10 x 10 ml) 100 test special waste collection point.
Mean (U/L) 433 923
REF: 283 003 ( 9 x 20 ml) 180 test S57: use appropriate container to avoid environmental
REF: 283 004 ( 4 x 60 ml) 240 test
REF: 283 005 ( 5 x 20 ml) 100 test CV% 1.57 0.71 S61: avoid release in environment. refer to special instructions/
safety data sheets.
Or prepare the working solution according to the number of tests
Intended Use required by mixing 4 volumes of reagent 1 (R1) and 1 volume of Run to run (Reproducibility)
Spectrum Diagnostics liquizyme LDH reagent is intended for the in- References
reagent 2 (R2),e.g. 400 ml R1 +100 ml R2. Level 1 Level 2
vitro quantitative, diagnostic determination of LDH in human serum 1. Dito WR. Lactate dehydrogenase: A brief review. In: Griffiths
on both automated and manual systems. n 20 20
Reagent Storage and Stability JC, ed. Clinical Enzymology. New York :masson publishing
All reagents are stable until expiration date stated on label when Mean (U/L) 439 935 USA; 1979:18
Background o
stored refrigerated at 2 - 8 C. SD 7.1 6.71
The lactate dehydrogenase (LDH) enzyme is widely distributed in o 2. Kachmar JF, Moss DW: Enzymes. In Fundamentals of clinical
Working solution is stable for 2 months at 2 – 8 C or 1 week
heart, liver, muscle, and kidney. LDH catalyzes the conversion of o
CV% 1.62 0.79 chemistry. NW Tietz, editor, saunders, philadelphia, 1976 pp
at 15 -25 C.
lactate to pyruvate. The enzyme is a tetrameric protein and gives 652-6603.
rise to five isoenzymes. Heart, kidney, brain and erythrocytes have Deterioration 3. Van der heiden C, B Ais, Gerh Ardt W,Rosallsis. Approved
the highest proportion of LD-1 and LD-2. Liver and skeletal muscle Do not use liquizyme LDH reagent if it is turbid or if the absorbance Methods Comparison recommendation on IFCC methods for the measurement of
have highest percentage of LD-5. LDH is significantly increased of the working reagent is less than 1.0 at 340 nm. A comparison between Spectrum Diagnostics LDH reagent and catalytic concentration of enzymes. Part 8. IFCC method for
during myocardial infarction. A maximum value is reached 48 Failure to recover control values within the assigned range may a commercial reagent of the same methodology was performed on LDH.Eur J Clinical Chem Clin Biochem. 1994;32:639-655
hours after the onset of manifestation and persists up to 10 days be an indication of reagent deterioration. 20 human sera. A correlation of 0.967 was obtained.
.Elevated serum levels of LDH have also been observed in patients 4. young DS.Effects of drugs on clinical laboratory tests. AACC
with megaloblastic anemia, disseminated carcinoma, leukemia, and Sensitivity press, Washington D.C., 1990
trauma. Mild increases in LDH activity has been reported in cases Use nonhaemolyzed serum. Heparin is the only acceptable When run as recommended, the minimum detection limit of this 5. Zimmerman HJ, henery JB: Clinical enzymology. In: Clinical
of haemolytic anemia, muscular dystrophy, pulmonary infarction, anticoagulant. Sodium citrate and EDTA have an inhibitor effect and assay is 10 U/L. diagnosis and management by laboratory methods, 16 th., JB
hepatitis, nephrotic syndrome, and cirrhosis. must not be used.The biological half-life of LDH in serum is 10 - 54 Henery, editor, saunders, philadelphia,1979, pp 365-368.
hours. Linearity
Method o
Stability: 6 weeks at 4 – 8 C ; 4 days at 20 – 25 C
o
The reaction is linear up to LDH concentration of 1200 U/L;
Kinetic ultraviolet method. Freezing of the samples is not recommended. specimens showing higher concentration should be diluted 1+5 with
physiological saline and repeat the assay (result×6).
Assay Principle System Parameters ORDERING INFORMATION
LDH catalyzes the reaction between pyruvate and NADH to Wavelength 340 nm (334 – 365 nm) Interfering substances
produce NAD+ and L-Lactate: Optical path 1 cm Serum, plasma CATALOG NO QUANTITY
Assay type Kinetic 283 001 4 x 20 ml
LDH Haemolysis
Pyruvate + NADH + H+ L- Lactate + NAD+ Direction decrease 283 002 10 x 10 ml
Sample : Reagent Ratio 1 : 50 Erythrocyte contamination elevates results significantly since LDH 283 003 9 x 20 ml
The initial rate of the NADH oxidation is directly proportional to the e.g.: Reagent volume 1 ml activities in erythrocytes are 150 times higher than those in normal 283 004 4 x 60 ml
catalytic LDH activity. It is determined by measuring the decrease in Sample volume 20 ml sera. 283 005 5 x 20 ml
o
absorbance at 340 nm. Temperature 37 C
Equilibration time 30 seconds. Icterus
Reagents Read time 1 to 3 minutes No significant interference.
Reagent 1 (R1 Buffer) Reagent Blank Limits Low 1.00 AU Lipemia
Phosphate buffer (pH 7.5) 50 mmol/L High 2.5 AU Lipemic specimens may cause high absorbance flagging. Diluted
Pyruvate 3.0 mmol/L Sensitivity 10 U/L sample may be recommended.
Sodium Azide 8.0 mmol/L Linearity 1200 U/L
Anticoagulants
Reagent 2 (R2 Coenzyme) Procedure EDTA and citrate may inhibit the reaction.
NADH > 0.18 mmol/L o
Pipette into cuvette (37 C): Macro Semi-Micro
Sodium azide 8.0 mmol/L Expected values (at 37 oC)
Working 1 ml 500 ml
solution Adults at 240-480 U/L (4.0- 8.0 mkat/L)
For further information, refer to the Lactate dehydrogenase reagent
material safety data sheet. Specimen 20 ml 10 ml Children (7-12 Years)
Female : < 580 U/L (< 9.65 mkat/L)
Precautions and Warnings Mix, read initial absorbance after 30 secnods. and start timer Male : < 764 U/L (< 12.7 mkat/L)
Do not ingest or inhalate. In case of contact with eyes or skin; rinse simultaneously. Read again after 1, 2 and 3 minutes. Determine the Premature : < 1103 U/L (< 18.4 mkat/L)
immediately with plenty of soap and water. In case of severe injuries; mean absorbance change per minute (DA/min).
seek medical advice immediately. Calculate for temperature conversion factor of
o o
0.5 (37 25 C) and 0.67 (37 30 C).
Calculation
Both reagents (R1) and (R2) contain sodium azide which may react To calculate the LDH activity use the following formula
with copper or lead plumbing. U/L = 8095 x DA 340 nm/min.
U/L = 15000 x DA 365 nm/min. Spectrum Diagnostics does not interpret the results of a
Reagent Preparation clinical laboratory procedure ; interpretation of the results is
REF:283 001: add 4 ml from R2 to one bottle of R1; mix gently. considered the responsibility of qualified medical personnel.
Quality Control
REF:283 002: add 2 ml from R2 to one bottle of R1; mix gently. All indications of clinical significance are supported by
REF:283 003: add 4 ml from R2 to one bottle of R1; mix gently. literature references.
be analyzed with each run.
REF:283004:add 12 ml from R2 to one bottle of R1; mix gently.
REF:283 005: add 4 ml from R2 to one bottle of R1; mix gently.
112 113
Lactate dehydrogenase SYMBOLS IN PRODUCT LABELLING
Run to run (Reproducibility) Waste Disposal
Level 1 Level 2
(LDH)-Liquizyme (1+1) EC REP Authorised Representative
Use by/Expiration Date n 20 20
E.C.1.1.1.27. LOT
REF
Catalogue Number
for use
Mean (U/L) 439 935
S57:
CV% 1.62 0.79 S61: avoid release in environment. refer to special instructions/
REF: 279 002 (4 x 25 ml) 100 test
safety data sheets.
Deterioration
Intended Use Do not use liquizyme LDH reagent if it is turbid or if the absorbance
Spectrum Diagnostics liquizyme LDH reagent is intended for the in- Methods Comparison Reference
of the working reagent is less than 1.0 at 340 nm. Failure to recover A comparison between Spectrum Diagnostics LDH reagent and a
vitro quantitative, diagnostic determination of LDH in human serum control values within the assigned range may be an indication of 1. Dito WR. Lactate dehydrogenase :A brief review. In : Griffiths
on both automated and manual systems. commercial reagent of the same methodology was performed on 20
reagent deterioration. JC, ed. Clinical Enzymology. New York :masson publishing
human sera. A correlation of 0.967 was obtained.
USA; 1979:18.
Background Specimen Collection and Preservation
The lactate dehydrogenase (LDH) enzyme is widely distributed in Sensitivity 2. Kachmar JF , Moss DW: Enzymes. In Fundamentals of clinical
Use nonhemolyzed serum. Heparin is the only acceptable When run as recommended, the minimum detection limit of this chemistry. NW Tietz, editor, saunders, philadelphia, 1976 pp
heart, liver, muscle, and kidney. LDH catalyzes the conversion of anticoagulant. Sodium citrate and EDTA have an inhibitor effect and
lactate to pyruvate. The enzyme is a tetrameric protein and gives assay is 10 U/L. 652- 6603.
must not be used.The biological half-life of LDH in serum is 10 - 54
rise to five isoenzymes. Heart, kidney, brain and erythrocytes have hours. 3. Van der heiden C, B AIS, Gerh Ardt W,Rosallsis. Approved
the highest proportion of LD-1 and LD-2. Liver and skeletal muscle o o Linearity
Stability: 6 weeks at 4 – 8 C ; 4 days at 20 – 25 C recommendation on IFCC methods for the measurement of
have highest percentage of LD-5. LDH is significantly increased The reaction is linear up to LDH concentration of 1200 U/L;
Freezing of the samples is not recommended. catalytic concentration of enzymes. Part 8. IFCC method for
during myocardial infarction. A maximum value is reached 48 specimens showing higher concentration should be diluted 1+5 with
LDH.Eur J Clinical Chem Clin Biochem . 1994;32:639-655.
hours after the onset of manifestation and persists up to 10 days physiological saline and repeat the assay (result×6).
System Parameters 4. young DS. Effects of drugs on clinical laboratory tests. AACC
.Elevated serum levels of LDH have also been observed in patients Wavelength 340 nm (334 – 365 nm)
with megaloblastic anemia, disseminated carcinoma, leukemia, and Interfering substances press, Washington D.C., 1990.
Optical path 1 cm
trauma. Mild increases in LDH activity has been reported in cases Serum, plasma
Assay type Kinetic 5. Zimmerman HJ, Henery JB : Clinical enzymology. In Clinical
of haemolytic anemia, muscular dystrophy, pulmonary infarction, Direction decrease diagnosis and management by laboratory methods, 16 th ed.,
hepatitis, nepherotic syndrome, and cirrhosis. Haemolysis
Sample : Reagent Ratio 1 : 50 JB henery, editor, saunders, philadeLphia,1979, pp 365-368.
Erythrocyte contamination elevates results significantly since LDH
Method activities in erythrocytes are 150 times higher than those in normal
Sample volume 20 ml
Kinetic ultraviolet method. o sera.
Temperature 37 C
Equilibration time 30 seconds.
Assay Principle Icterus
Read time 1 to 3 minutes ORDERING INFORMATION
LDH catalyzes the reaction between pyruvate and NADH to No significant interference.
produce NAD and L-Lactate: Reagent Blank Limits Low 1.00 AU CATALOG NO QUANTITY
Lipemia
High 2.5 AU 279 001 2 x 25 ml
LDH Lipemic specimens may cause high absorbance flagging.Diluted
Pyruvate + NADH + H+ L- Lactate + NAD+ Sensitivity 10 U/L 279 002 4 x 25 ml
sample may be recommended.
Linearity 1200 U/L
The initial rate of the NADH oxidation is directly proportional to the Anticoagulants
catalytic LDH activity. It is determined by measuring the decrease in Procedure
EDTA and citrate may inhibit the reaction.
absorbance at 340 nm. Pipette into cuvette ( 37 C )
o
Working 1 ml ( or add 0.5 ml R1 + 0.5 ml R2 ) Expected value (at 37 oC)

Reagents
solution Adults 240-480 U/L (4.0- 8.0 mkat/L)
Phosphate buffer (pH 7.5) 50 mmol/L Specimen 20 ml Children (7-12 Years)
Pyruvate 3.0 mmol/L Female : < 580 U/L (< 9.65 mkat/L)
Sodium Azide 8.0 mmol/L Mix, read initial absorbance after 30 secnods. and start timer
simultaneously. Read again after 1, 2 and 3 minutes. Determine the Male : < 764 U/L (< 12.7 mkat/L)
Reagent 2 (R2 Enzyme) Premature : < 1103 U/L (< 18.4 mkat/L)
NADH > 0.06 mmol/L mean absorbance change per minute (DA/min).
Sodium azide 8.0 mmol/L Calculate for temperature conversion factor of
o o
Calculation 0.5 (37 25 C) and 0.67 (37 30 C).
For further information, refer to the Lactate dehydrogenase reagent To calculate the LDH activity use the following formula
material safety data sheet. U/L = 8095 x DA 340 nm/min.
Precautions and Warnings Quality Control Spectrum diagnostics does not interpret the results of a
Normal & abnormal control serum of known concentrations should clinical laboratory procedure; interpretation of the results is
Do not ingest or inhalate. In case of contact with eyes or skin; rinse be analyzed with each run. considered the responsibility of qualified medical personnel.
immediately with plenty of soap and water. In case of severe injuries; All indications of clinical significance are supported by
seek medical advice immediately. Performance Characterstics literature references.
Precision
Both reagents (R1) and (R2) contain sodium azide which may react
with copper or lead plumbing.
Analytical Range:
Reagent Preparation, Storage and Stability Level 1 Level 2 10 - 1200 U/L.
All reagents are stable until expiration date stated on label when n 20 20
o
stored refrigerated at 2 - 8 C.
Working solution can be prepared by adding equal volumes from Mean (U/L) 433 923
R1 and R2 . SD 6.8 6.64
o o CV% 1.57 0.71
Stability: 2 months at 2 – 8 C or 1 week at 15 – 25 C.
114 115
LIPASE-LS SYMBOLS IN PRODUCT LABELLING
Analytical range
3 U/l - 300 U/l.
COLORIMETRIC (DGMRE) EC REP Authorised Representative
Use by/Expiration Date Precision
Within run Mean [U/l] SD [U/l] CV [%]
REF:281 001 40 test REF Catalogue Number for use n = 40
Sample 1 13,4 0,24 1,81
Sample 2 58,9 0,60 1,01
Intended use SAMPLES Sample 3 103 1,50 1,45
Spectrum diagnostics Lipase-LS reagent is intended for in-vitro Serum free of hemolysis, Heparin plasma.
quantitative determination of Lipase in human serum, heparinized Stability : 24 hrs at 15 - 25 °C Beween run Mean [U/l] SD [U/l] CV [%]
or EDTA plasma. 5 days at 2 - 8 °C n = 40
1 year at -20 °C
Background Sample 1 13,4 0,24 1,81
Pancreatic lipase in serum is closely associated with pancreatic Sample 2 58,9 0,49 0,82
PROCEDURE
diseases. The activity of this enzyme has been measured as an Sample 3 103 0,65 0,63
This reagent can be used manually (see method below) and on
important marker for diagnosing pancreatic diseases and the most analyzers. Applications are available on request.
associated monitoring of therapeutic effects.Pancreatic lipase Correlation
test kits currently available include a turbidimetric method using Wavelength 580 nm, Hg 578 nm A comparative study has been performed between the Spectrum
triglyceride as substrate and a colorimetric method using synthetic Cuvette 1 cm method and another commercial reagent on 67 human serum
substrates. Temperature 37 °C samples.
Measure Against air
METHOD The parameters of linear regression are as follows:
Colorimetric Test, Kinetic y = 0,96 x – 1,15 U/l r = 0,999
Reagent Sample /
Blank Calibrator
PRINCIPLE INTERFERING SUBSTANCES
Lipase catalyzes the following reaction : Sample /Calibrator ------ 10 µl - Ascorbic Acid: no interference up to 30 mg/dL
1,2-o-Dilauryl-rac-glycero-3-glutaric acid-(6-methylresorufin)-ester - Bilirubin: no interference up to 60 mg/dL
dist. water 10 µl ------
- Hemoglobin: no interference up to 500 mg/dL
Lipase /Colipase
Reagent 1 500 µl 500 µl - Triglycerides: no interference up to 1000 mg/dL
1,2-o-Dilauryl-rac-glycerol + Glutaric acid-(6-methylresorufin)-ester
Mix carefully (do not shake), incubate 1 to 5 min, then add R2 to
start the reaction : Refrences
Cleavage 1. Lorentz K. Lipase. In: Thomas L, editor. Clinical laboratory
Glutaric acid -(6-methylresorufin)-ester Glutaric acid + Reagent 2 125 µl 125 µl
Methylresorufin diagnostics.1st ed. Frankfurt: TH-Books Verlags-gesellschaft;
Mix carefully (do not shake), incubate for 2 min at 37 °C, read 1998. p. 95-7.
absorbance and start stopwatch. After 1 min and after 2 min read
A synthetic substrate (DGMRE) is split by Lipase to yield the colored 2. Moss DW, Henderson AR. Digestive enzymes of pancreatic
absorbance again.
final product Methylresorufin. The increasing absorbance of the red origin. In: Burtis CA, Ashwood ER, editors. Tietz Textbook
Methylresorufin is measured photometrically . of Clinical Chemistry. 3rd ed. Philadelphia: W.B Saunders
The reaction is highly specific on the human enzyme. CALCULATION
Company; 1999. p. 689-708.
DA/min = [ A/minSample / Calibrator ] – [ A/minReagent Blank ] 3. Tietz N, Shuey DF. Lipase in serum – the elusive enzyme: an
REAGENTS
Reagents Composition (concentrations in the test): overview. Clin Chem 1993;39:746-56.
DA/minSample
Lipase (U/l) = x Conc.Calibrator 4. Lott J, Patel ST, Sawhney AK, Kazmierczak SC, Love JE.
Reagent 1: DA/minCalibrator Assays of serum lipase: analytical and clinical considerations.
Goods Buffer pH 8,0 40 mmol/l Clin Chem 1986;32:1290-1302.
Taurodesoxycholate 3,4 mmol/l
Desoxycholate 2,6 mmol/l
Expected Values 5. Leybold A, Junge W. Importance of colipase for the
Calciumchloride 12 mmol/l < 60 U/l measurement of serum lipase activity. Adv Clin Enzymol
Colipase 1 mg/l 1986;4:60-7.
Note: It is recommended for each laboratory to establish and
maintain its own reference values. The given data are only an 6. Borgström B. The action of bile salts and other detergents on
Reagent 2:
indication. pancreatic lipase and the interaction with colipase. Biochimica
Tartrate Buffer pH 4,0 1,5 mmol/l
et Biophysika Acta 1977;488:381-91.
Taurodesoxycholate 3,4 mmol/l
DGMRE 0,13 mmol/l CALIBRATOR & CONTROLS 7. Gargouri Y, Julien R, Bois A, Verger R, Sarda L. Studies on
Coemulgator For the calibration of automated analyzers Spectrum Multicalibrator the detergent inhibition of pancreatic lipase activity. J of Lipid
is recommended, for quality control use Spectrum normal and Research 1983;24:1336-42.
Precautions abnormal controls.
- For in vitro diagnostic use only.
Sensitivity
The detection limit is equal to 3 U/l. ORDERING INFORMATION
Stability
When stored at 2-8° C and protected from light, the reagents are CATALOG NO QUANTITY
stable up to the expiry date printed on the labels. Linearity 281 001 40 test
The reagent is linear up to 300 U/l.
Preparation and stability of Working reagents If this level is passed, repeat the test using serum diluted 1 +1 with
The reagents are ready to use. sodium chloride solution (9 g/L). Multiply result by 2.
Stability : 3 months at 2-8°C, if contamination is avoided
116 117
iii-Electrolytes
118 119
Ammonia II SYMBOLS IN PRODUCT LABELLING
Mix, and immediately read the absorbance A1 of the standard or
specimen against reagent blank. Exactly 5 minutes. later, read
Anticoagulants
Fluoride, citrate, and heparin must not be used.
(GLDH - UV - Test) EC REP Authorised Representative
absorbance A2 of standard or specimen.
Drugs
LOT Batch Code/Lot number CAUTION. Consult instructions Calculation Sodium cefoxitin causes artificially high ammonia values at the
REF: 333 001 100 tests REF Catalogue Number for use tested drug level.
Consult instructions for use Manufactured by A1 – A2 = DAspecimen or DAstandard .
R1a : 5 x 18 ml
R1b : 1 x 11 ml Expected Values
R2 : 1 x 12 ml EDTA plasma
Concentration of ammonia in Plasma: Adults
Females 19- 87 mg/dL (11-51 mmol/L)
Reagent preparation Males 27-102 mg/dL (16-60 mmol/L)
Intended Use DAspecimen
Prepare R1 Mixture as following: Ammonia (mg/dl) = x 521 Children < 81.5 mg/dL ( < 48 mmol/L)
Spectrum Diagnostics ammonia II reagent is intended for the in-vitro REF: 221 001:add 2 ml from R1b to one bottle of R1a;mix gently DAstandard
quantitative, diagnostic determination of ammonia in human plasma Neonates (1- 6 days) < 228 mg/dL (< 134 mmol/L)
Or prepare the working solution according to the number of tests
on both automated and manual systems. required by mixing 9 volumes of reagent 1a (R1a) and 1volume of Quality Control Spectrum Diagnostics does not interpret the results of a
reagent 1b (R1b); eg. 900 ml R1a +100 ml R1b. Normal & abnormal commercial control of known concentrations
Background clinical laboratory procedure; interpretation of the results is
should be analyzed with each run. considered the responsibility of qualified medical personnel.
Ammonia enters the body in nitrogen-containing foods via the Reagent Storage and Stability
gastrointestinal tract and is excreted largely as urea in urine and All reagents are stable until expiration date stated on label when All indications of clinical significance are supported by
Performance Characteristics literature references.
as bacterial protein in feces. Ammonia, the end product of nitrogen o
stored refrigerated at 2 - 8 C.
metabolism is absorbed into the portal venous blood and, after o Precision
R1 Mixture is stable for 1 month at 2– 8 C.
passing through the liver enters the systemic circulation. Normally
about half the ammonia is extracted from the body by the skeletal Within run (Repeatiblity) Analytical Range
Deterioration 9 – 1700 mg/dL.
muscle and about 16 % by the liver and brain. Clinically, the ext- Do not use Ammonia II reagent if it is turbid or if the absorbance of Level 1 Level 2
raction of ammonia by individual organs has different implications. the working reagent is less than 1.0 at 340 nm. Failure to recover
The hepatic conversion of ammonia to urea represents the primary n 20 20 Waste Disposal
mechanism of eliminating ammonia from the body. Conversely, Mean (mg/dL) 1.8 3.5 This product is made to be used in professional laboratories.
the excessive uptake of ammonia by the brain results in ammonia Please consult local regulations for a correct waste disposal.
SD 0.04 0.06 S56: dispose of this material and its container at hazardous or
intoxication, increased intracranial pressure and hepatic enceph-
Specimen Collection and Preservation special waste collection point.
alopathy. Hyperammonemia in infants may be due to inherited CV% 2.3 1.3
EDTA is the only acceptable anticoagulant because it reduces red cell S57: use appropriate container to avoid environmental
deficiencies of the urea cycle enzymes or acquired through acute
ammonia production. Other anticoagulants produce sponteniously contamination.
(as in Reye’s syndrome) or chronic (as in cirrhosis) liver disease.
high results. Collect blood from stasis-free vein of fasting patient. S61: avoid release in environment. refer to special instructions/
run to run (Reproducibility)
Smoking should be avoided prior to sample. Tubes should be filled safety data sheets.
Method completely and kept tightly stoppered at all times. Place immediately Level 1 Level 2
Kinetic enzymatic method with glutamate dehydrogenase. o
on ice and centrifuge, preferrable at 4 C. Perform analysis within 30
n 20 20 References
minutes of venipuncture.
Assay Principle Mean (mg/dL) 1.8 3.5 1. Burtis CA, Ashwood ER, eds. Tietz fundamentals of Clinical
∝ – ketoglutarate reacts with ammonium ions in presence of Note: avoid contamination of samples by ammonia from smoking SD 0.07 0.14 Chemistry. 4 th ed. Philadelphia: WB saunders: 1996:755.
glutamate dehydrogenase and the coenzyme NADH to produce or traffic in laboratory or patient’s room, glassware, or water. One
known source of spontaneous ammonia formation is an increased CV% 3.4 4.1 2. Dewan JG: the L[+]-glutamic dehydrogenase of animal tissue
L-glutamate and NAD+
Biochem J 32”1378,1938.
∝-glutamyl-transferase activity leading to decomposition of
NH4+ + ∝-KG L-glutamate glutamine. 3. Diamond EG: Inhibitory efect of heparin upon adenylic
GLDH +
+ o o
Stability: 15 minutes. at 15 – 25 C; 2 hours at 4 – 8 C; Methods Comparison deaminase. J lab Clin Med 46:807,1955.
NADH NAD+ + H2O o A comparison between Spectrum Diagnostics Ammonia reagent and
3 weeks at -20 C
a commercial reagent of the same methodology was performed on 4. Howanitz JH, Howanitz PJ, Skrodzki CA, Iwanski JA.
The concentration of The NADP+ formed is directly proportional 20 human plasma. A correlation of 0.976 was obtained. Influences of specimen processing and storage condition on
System Parameters
to the ammonia concentration. It is determined by measuring the results for plasma ammonia. Clin Chem. 1984;30:906-908.
Wavelength 340 nm
decrease in absorbance at 340 nm. Optical path 1 cm Sensitivity 5. Olson JA, anfinsen CB: kinetic and equilibrium studies on
Assay type Fixed Rate When run as recommended, the minimum detection limit of this crystalline L-glutamate acid dehydrogenase. J Biol Chem
Reagents Direction Decrease assay is 9.0 mg/dL. 202:841, 1953
Standard ammonia (ST) First read time 2 seconds
521 mg/dL 307 mmol/L Delay time 300 seconds Linearity 6. Vananken HC, Scphorst ME. A kinetic determination of
last read time 5 Minutes The reaction is linear up to ammonia concentration of 1700 mg/dL. ammonia in plasma. Clin Chem Acta.1974;56:151-157.
o
Reagent R1a Buffer Temperature 37 C 7. Young DS:et al. Clin Chem. 1975; 21.
Tris buffer (pH 8.0) 50 mmol/L Zero adjustment Reagent blank Interfering Substances
GLDH > 6 kU/L Reagent Blank Limits Low 1.00 AU plasma
LDH > 2 kU/L High 2.0 AU
EDTA 5.3 mmol/L Sensitivity 9 mg/dL (5.3 mmol/L) Haemolysis
Linearity 1700 mg/dL (1000 mmoL/L) Avoid haemolyzed specimen since RBCs contain three times the
Reagent R1b ORDERING INFORMATION
ammonia content of plasma.
Coenzyme ( NADH) 0.18 mmol/L Procedure CATALOG NO QUANTITY
Icterus 333 001 100 Tests
Reagent R2 Standard Specimen Reagent blank
Bilirubin levels higher than 30 mg/dL increase the ammonia
∝ – ketoglutarate 18 mmol/L R1 mixture 1ml 1ml 1ml concentration significantly.
Standard 100 ml ----- -----
For further information, refer to the Ammonia reagent material safety ∝-globulin
data sheet. Specimen ----- 100 ml -----
Elevated ∝-globulin levels (more than 3 g/dL) may increase the
Dist. H2O ----- ----- 100 ml apparent ammonia concentration values.
Precautions and Warnings o
Incubate for 3 minutes at 37 C, Then add
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Lipemia
immediately with plenty of soap and water. In case of severe injuries; R2 100 ml 100 ml 100 ml Lipemic samples should be centrifuged and the analysis performed
seek medical advice immediately. on the supernatent.
120 121
Ammonia – Liquizyme SYMBOLS IN PRODUCT LABELLING
Calculation
A1 – A2 = Aspecimen or Astandard or Aspecimen blank.
Lipemia
Lipemic samples should be centrifuged and the analysis performed
on the supernatent.
Aspecimen Final = Aspecimen – Aspecimen blank.
LOT Batch Code/Lot number CAUTION. Consult instructions Anticoagulants
REF: 220 001 (2 x 20 ml) Fluoride, citrate, and heparin must not be used.
REF: 220 002 (5 x 20 ml) REF Catalogue Number for use
Drugs
Consult instructions for use Manufactured by Concentration of ammonia in serum: Sodium cefoxitin causes artificially high ammonia values at the
tested drug level.
Intended Use Reagent Storage and Stability Aspecimen Final
Spectrum Diagnostics liquizyme ammonia single reagent is intended All reagents are stable until expiration date stated on label when Ammonia (mg/dl) = x 521 Expected Values
Astandard
for the in-vitro quantitative, diagnostic determination of ammonia in o
stored refrigerated at 2 - 8 C. EDTA plasma
human plasma on both automated and manual systems. Adults
Deterioration Quality Control Females 19- 87 mg/dL (11-51 mmol/L)
Background Do not use liquizyme Ammonia reagent if it is turbid or if the Normal & abnormal commercial control serum of known Males 27-102 mg/dL (16-60 mmol/L)
Ammonia enters the body in nitrogen-containing foods via the absorbance of the working reagent is less than 1.0 at 340 nm. concentrations should be analyzed with each run. Children < 81.5 mg/dL ( < 48 mmol/L)
gastrointestinal tract and is excreted largely as urea in urine and Failure to recover control values within the assigned range may be Neonates (1- 6 days) < 228 mg/dL (< 134 mmol/L)
as bacterial protein in feces. Ammonia, the end product of nitrogen an indication of reagent deterioration. Performance Characteristics
metabolism is absorbed into the portal venous blood and, after Precision
passing through the liver enters the systemic circulation. Normally Spectrum Diagnostics does not interpret the results of a
Specimen Collection and Preservation clinical laboratory procedure; interpretation of the results is
about half the ammonia is extracted from the body by the skeletal EDTA is the only acceptable anticoagulant because it reduces red cell Within run (Repeatiblity) considered the responsibility of qualified medical personnel.
muscle and about 16 % by the liver and brain. Clinically, the ext- ammonia production. Other anticoagulants produce sponteniously
raction of ammonia by individual organs has different implications. Level 1 Level 2 All indications of clinical significance are supported by
high results. Collect blood from stasis-free vein of fasting patient. literature references.
The hepatic conversion of ammonia to urea represents the primary Smoking should be avoided prior to sample. Tubes should be filled n 20 20
mechanism of eliminating ammonia from the body. Conversely, completely and kept tightly stoppered at all times. Place immediately
the excessive uptake of ammonia by the brain results in ammonia o Mean (mg/dL) 1.8 3.5
on ice and centrifuge, preferrable at 4 C. Perform analysis within 30 Analytical Range
intoxication, increased intracranial pressure and hepatic enceph- minutes of venipuncture. SD 0.04 0.06 9 – 1700 mg/dL.
alopathy. Hyperammonemia in infants may be due to inherited
deficiencies of the urea cycle enzymes or acquired through acute CV% 2.3 1.3
Note: avoid contamination of samples by ammonia from smoking Waste Disposal
(as in Reye’s syndrome) or chronic (as in cirrhosis) liver disease. or traffic in laboratory or patient’s room, glassware, or water. One This product is made to be used in professional laboratories.
known source of spontaneous ammonia formation is an increased run to run (Reproducibility) Please consult local regulations for a correct waste disposal.
Method ∝-glutamyl-transferase activity leading to decomposition of S56: dispose of this material and its container at hazardous or
Kinetic enzymatic method with glutamate dehydrogenase. Level 1 Level 2
glutamine. special waste collection point.
o o
Stability: 15 minutes. at 15 – 25 C; 2 hours at 4 – 8 C; n 20 20 S57: use appropriate container to avoid environmental
Assay Principle 3 weeks at -20 C
o
contamination.
Mean (mg/dL) 1.8 3.5
∝ – ketoglutarate reacts with ammonium ions in presence of S61: avoid release in environment. refer to special instructions/
glutamate dehydrogenase and the coenzyme NADPH to produce System Parameters SD 0.07 0.14 safety data sheets.
L-glutamate and NADP+ Wavelength 340 nm CV% 3.4 4.1
Optical path 1 cm References
NH4 + ∝-KG
+ L-glutamate Assay type Fixed Rate
GLDH +
+ Direction Decrease 1. Burtis CA, Ashwood ER, eds. Tietz fundamentals of Clinical
NADPH NADP+ + H2O Methods Comparison Chemistry. 4 th ed. Philadelphia: WB saunders: 1996:755.
Sample : Reagent Ratio 1 : 10 A comparison between Spectrum Diagnostics Ammonia single
e.g.: Reagent volume 1 ml reagent and a commercial reagent of the same methodology was 2. Dewan JG: the L[+]-glutamic dehydrogenase of animal tissue.
The concentration of The NADP+ formed is directly proportional Sample volume 100 ml Biochem J 32”1378,1938.
performed on 20 human sera. A correlation of 0.978 was obtained.
to the ammonia concentration. It is determined by measuring the First read time 30 seconds
decrease in absorbance at 340 nm. Delay time 150 seconds 3. Diamond EG: Inhibitory efect of heparin upon adenylic
Sensitivity deaminase. J lab Clin Med 46:807,1955.
last read time 180 seconds
Reagents Temperature 37 C
o
4. Howanitz JH, Howanitz PJ, Skrodzki CA, Iwanski JA. Influences
assay is 9.0 mg/dL.
Standard ammonia (ST) Zero adjustment Against Air of specimen processing and storage condition on results for
521 mg/dL 307 mmol/L Reagent Blank Limits Low 1.00 AU
Linearity plasma ammonia. Clin Chem. 1984;30:906-908.
High 2.0 AU
Reagent (R) Sensitivity 9 mg/dL (5.3 mmol/L) The reaction is linear up to ammonia concentration of 1700 mg/dL. 5. Olson JA, anfinsen CB: kinetic and equilibrium studies on
Bicine buffer (pH 8.5) 100 mmol/L Linearity 1700 mg/dL (1000 mmoL/L) crystalline L-glutamate acid dehydrogenase. J Biol Chem
Interfering Substances 202:841, 1953
∝ – ketoglutarate 7.5 mmol/L
Sodium Azide 0.05% Procedure plasma
6. Vananken HC, Scphorst ME. A kinetic determination of
GLDH (microbial) 500 Ku/L Reagent blank Standard Specimen ammonia in plasma. Clin Chem Acta.1974;56:151-157.
NADPH 0.2 mmol/L Haemolysis
Sodium Azide 8 mmol/L Reagent (R) 1.0 ml 1.0 ml 1.0 ml Avoid haemolyzed specimen since RBCs contain three times the 7. Young DS:et al. Clin Chem. 1975; 21.
ammonia content of plasma.
Standard --------- 100 ml ---------
For further information, refer to the Ammonia reagent material safety
data sheet. Specimen --------- --------- 100 ml Icterus
Bilirubin levels higher than 30 mg/dL increase the ammonia ORDERING INFORMATION
Mix, and after 30 seconds. read the absorbance A1 of the standard concentration significantly.
Precautions and Warnings CATALOG NO QUANTITY
Do not ingest or inhalate. In case of contact with eyes or skin; rinse or specimen or specimen blank. Exactly 2.5 minutes. later, read
absorbance A2 of standard or specimen or specimen blank. ∝-globulin 220 001 2 x 20 ml
immediately with plenty of soap and water. In case of severe injuries; 220 002 5 x 20 ml
seek medical advice immediately. Elevated µ-globulin levels (more than 3 g/dL) may increase the
o
*Note: It is recommended to Incubate reagent at 37 C for 3 minutes. apparent ammonia concentration values.
The reagent (R) contain sodium azide which may react with copper then add 100 ml of the serum or standard to each 1 ml and complete
or lead plumbing. the procedure as above.
Reagent preparation
Spectrum Ammonia single reagent is supplied ready-to-use and
stable up to the expiry date labeled on the bottles Once opened,
the opened vial is stable for 3 months at the specified temperature.
122 123
Calcium
o
Mix and incubate for 5 minutes at 20 - 25 C. Measure absorbance Expected values
SYMBOLS IN PRODUCT LABELLING
of specimen (Aspecimen) and standard (Astandard) against reagent Serum, plasma
O-CPC EC REP Authorised Representative
blank. Adults
20 - 50 years 8.8-10.2 mg/dl (2.20-2.55 mmol/L)
LOT Batch Code/Lot number for use Calculation (Aspecimen) >50 years 8.4- 9.7 mg/dl (2.09-2.42 mmol/L)
REF: 226 001 (2 x 30 ml) 60 test REF Catalogue Number Manufactured by Serum calcium concentration (mg/dL) = x 10
Consult instructions for use (Xi) - Irritant (Astandard) Children
REF: 226 002 (2 x100 ml) 200 test
4 -18years 9.2-11.0 mg/dl (2.30-2.75 mmol/L)
REF: 226 003 (4 x100 ml) 400 test Temperature Limitation (Aspecimen)
Urine calcium (mg/24 hrs)= x10 x10*x 2**x V*** >4 weeks 7.2-11.2 mg/dl (1.80-2.8 mmol/L)
REF: 226 004 (2 x 50 ml) 100 test
(Astandard) Urine (24 h)
Females <250 mg/day (<6.25 mmol/day)
Intended Use Reagent Preparation, Storage and Stability * The factor “10” converts mg/dl to mg/litre Males <300 mg/day (<7.5 mmol/day)
Spectrum Diagnostics calcium reagent is intended for the in-vitro Spectrum Calcium reagents are supplied ready-to-use and stable ** The factor “2” represents the dilution factor Childern <6 mg/Kg/day (<0.15 mmol/day)
quantitative, diagnostic determination of calcium in human serum on up to the expiry date labeled on the bottles when stored sealed at *** “V” represents the 24-hour urine volume in litres
o
both automated and manual systems. 15 – 25 C.
Quality Control Spectrum Diagnostics does not interpret the results of a
Background Deterioration Normal & abnormal control serum of known concentrations should clinical laboratory procedure;interpretation of the results is
Calcium is the fifth most common element in the body, most of Do not use the Spectrum Calcium reagents if turbid. Failure to be analyzed with each run. considered the responsibility of qualified medical personnel.
which (98 %) is present in the skeleton. One half of the remaining recover control values within the assigned range may be an All indications of clinical significance are supported by
calcium is found in extracellular fluid and the rest in tissues. Calcium indication of reagent deterioration. Performance Characterstics literature references.
has a crucial role in bone mineralization and is also vital for basic Precision
physio-logical processes such as blood coagulation, neuromuscular Specimen Collection and Preservation
conduction, and normal muscle tone. Calcium is constantly lost from Serum and plasma Within run (Repeatiblity) Analytical Range
the body through excretion in feces, urine and to a small extent in Use nonhemolyzed serum.Heparin is the only acceptable antico- 2 – 20 mg/dl (0.5-5 mmol/L).
agulant. No other anticoagulant can be used. Fresh serum collected Level 1 Level 2
sweat. The determination of serum calcium is useful for monitoring
myeloma, renal failure, acid base balance,and cirrhosis. Both in the fasting state is the perferred specimen. Serum or plasma n 20 20 Waste Disposal
serum and tissue calcium in the body are controlled by parathyroid should be separated from cells as soon as possible, because This product is made to be used in professional laboratories.
prolonged contact with the clot may cause lower calcium values. Mean (mg/dL) 9.58 13.97
hormone, calcitonin and vitamin D. Hypocalcemia may be observed Please consult local regulations for a correct waste disposal.
in hypoparathyrodism, steatorrhea, pancreatitis and nephrosis. Sera from patients receiving EDTA (treatment of hypercalcemia) SD 0.12 0.207 S56: dispose of this material and its container at hazardous or
Increased levels may be associated with multiple myeloma and are unsuitable for analysis, since EDTA will chelate the calcium and special waste collection point.
CV% 1.33 1.48
other neoplastic diseases. render it unavailable for reaction with O-cresolphthalein complexone. S57: use appropriate container to avoid environmental
The biological half-life of calcium in blood is few hours. contamination.
Method Run to run (Reproducibility) S61: avoid release in environment. refer to special instructions/
O-cresolphthaline complexone colorimetric method. Urine safety data sheets.
Specimens should be collected in acid washed bottles. 24 hour Level 1 Level 2
Assay Principle Specimens should be collected in containers containing 5 ml of n 20 20
Calcium ions react with O-cresolphthalein complexone (O-CPC) 6 mol/L HCl. If the specimen is collected without acid, the pH should References
be adjusted < 3 with 6 mol/L HCl. Dilute urine specimen 2 times with Mean (mg/dL) 9.62 14.15
under alkaline conditions to form a violet colored complex. 1. Barnett RN: A scheme for the comparison of quantitative
magnesium and iron. bidistilled water (1volume urine + 1volume distilled water) before SD 0.23 0.221 methods. AM J Clin Pathol 43: 562, 1965.
assay.
CV% 1.42 1.53
Alkaline pH 2. Fiereck EA: Appendix. Normal values. in:Fundamentals of
Ca2+ + O-CPC calcium-O-CPC complex o
Stability (serum): 7 days at 15 – 25 C; 3 weeks at 4 – 8 C;
o
clinical chemistry. NW Tietz, editor,Saunders, Philadelphia,
o
8 months at -20 C Methods Comparison p1208,1976.
The color intensity of the complex formed is directly proportional A comparison between Spectrum Diagnostics Calcium reagent and
to the calcium concentration. It is determined by measuring the Stability (urine): 2 days at 15 – 25 oC; 4 days at 4 – 8 C;
o
a commercial reagent of the same methodology was performed on 3. Kessler G, wolfman M: An automated procedure for the
increase in absorbance at 578 nm. o
3 weeks at -20 C 20 human sera. A correlation of 0.979 was obtained. simultaneous determination of calcium and phosphorus.Clin
Chem 10:686, 1964.
Reagents Stored serum or urine specimens must be mixed well prior to Sensitivity 4. Peters JP, Van Slyke, DD: Quantitative clinical chemistry, vol
analysis. When run as recommended, the minimum detection limit of this 2,williams and wilkins, Baltimor (MD),1932, p 760.
Standard Calcium (ST) assay is 2.0 mg/dL.
10 mg/dL 2.5 mmol/L System Parameters 5. Tietz NW: Blood gases and electrolytes. In:Fundamentals of
Wavelength 578 nm Linearity clinical chemistry, NW tietz, editor,Saunders, Philadelphia,
Reagent 1 (R1 Buffer) Optical path 1 cm The reaction is linear up to calcium concentration of 20 mg/dl. 176,pp 903, 908.
2-Amino-2-methyl-1-propanol (pH 10.5) 0.3 mol / L Assay type End-point Specimens showing higher concentration should be diluted 1+1 6. Young DS, Effects of drugs on clinical laboratory tests. AACC
Direction Increase using physiological saline and repeat the assay (result × 2). press, Washington, D.C.1990.
Reagent 2 (R2 Chromogen) Sample : Reagent Ratio 1 : 100
O-cresolphthalein complexone 0.16 mmol/L e.g.: Reagent volume 1 ml Interfering Substances:
8-hydroxyquinoline 7.0 mmol/L Sample volume 10 ml
o
Temperature 15 - 25 C Haemolysis
Irritant (Xi) Zero adjustment Reagent Blank avoid haemolysis.
R20/21/22 Harmful by inhalation, in contact with skin Reagent Blank Limits Low 0.00 AU ORDERING INFORMATION
and if swallowed. High 0.15 AU Icterus CATALOG NO QUANTITY
R38 Irritating to skin. Sensitivity 2 mg/dL (0.5 mmol/L) No significant interference.
R41 Risk of serious damage to eyes. 226 001 2 x 30 ml
Linearity 20 mg/dL (5 mmol/L)
S24/25 Avoid contact with skin and eyes. 226 002 2 x 100 ml
Lipemia 226 003 4 x 100 ml
S26 In case of contact with eyes, rinse immediately Procedure No significant interference.
with plenty of water and seek medical advice. 226 004 2 x 50 ml
For further information, refer to the Calcium reagent material safety Blank Standard Specimen
Anticoagulants
data sheet. Standard ------ 10 ml ------ complexing Anticoagulants such as citrate, oxalate and EDTA must
Specimen ------ ------ 10 ml be avoided.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Reagent 1 0.5 ml 0.5 ml 0.5 ml
Reagent 2 0.5 ml 0.5 ml 0.5 ml
124 125
Calcium SYMBOLS IN PRODUCT LABELLING
Precision
clinical laboratory procedure;interpretation of the results is
Arsenazo III (Single Reagent) EC REP Authorised Representative
REF: 227 001 (2 x 30 ml) 60 test LOT Batch Code/Lot number CAUTION. Consult instructions Level 1 Level 2 literature references.
REF: 227 002 (2 x100 ml) 200 test REF Catalogue Number for use
n 20 20
Mean (mg/dL) 9.58 13.97 Analytical Range
SD 0.12 0.207 2 – 20 mg/dl (0.5-5 mmol/L).
Intended Use Urine CV% 1.33 1.48 Waste Disposal
Spectrum Diagnostics calcium reagent is intended for the in-vitro Specimens should be collected in acid washed bottles. 24 hour
quantitative, diagnostic determination of calcium in human serum on Specimens should be collected in containers containing 5 ml of This product is made to be used in professional laboratories.
Run to run (Reproducibility) Please consult local regulations for a correct waste disposal.
both automated and manual systems. 6 mol/L HCl. If the specimen is collected without acid, the pH should
be adjusted < 3 with 6 mol/L HCl. Dilute urine specimen 2 times with Level 1 Level 2 S56: dispose of this material and its container at hazardous or
Background bidistilled water (1volume urine + 1volume distilled water) before special waste collection point.
n 20 20 S57: use appropriate container to avoid environmental
Calcium is the fifth most common element in the body, most of assay.
which (98 %) is present in the skeleton. One half of the remaining Mean (mg/dL) 9.62 14.15 contamination.
calcium is found in extracellular fluid and the rest in tissues. Calcium
o
Stability (serum): 7 days at 15 – 25 C; 3 weeks at 4 – 8 C;
o S61: avoid release in environment. refer to special instructions/
SD 0.23 0.221
has a crucial role in bone mineralization and is also vital for basic
o
8 months at -20 C safety data sheets.
CV% 1.42 1.53
physio-logical processes such as blood coagulation, neuromuscular
conduction, and normal muscle tone. Calcium is constantly lost from Stability (urine): 2 days at 15 – 25 oC; 4 days at 4 – 8 C;
o
References
the body through excretion in feces, urine and to a small extent in
o
3 weeks at -20 C Methods Comparison
A comparison between Spectrum Diagnostics Calcium reagent and 1. Barnett RN: A scheme for the comparison of quantitative
sweat. The determination of serum calcium is useful for monitoring methods. AM J Clin Pathol 43: 562, 1965.
myeloma, renal failure, acid base balance,and cirrhosis. Both Stored serum or urine specimens must be mixed well prior to a commercial reagent of the same methodology was performed on
serum and tissue calcium in the body are controlled by parathyroid analysis. 20 human sera. A correlation of 0.979 was obtained. 2. Fiereck EA: Appendix. Normal values. in:Fundamentals of
hormone, calcitonin and vitamin D. Hypocalcemia may be observed clinical chemistry. NW Tietz, editor,Saunders, Philadelphia,
in hypoparathyrodism, steatorrhea, pancreatitis and nephrosis. System Parameters Sensitivity p1208,1976.
Increased levels may be associated with multiple myeloma and Wavelength 650 nm (600 nm) When run as recommended, the minimum detection limit of this
assay is 2.0 mg/dL. 3. Kessler G, wolfman M: An automated procedure for the
other neoplastic diseases. Optical path 1 cm
simultaneous determination of calcium and phosphorus.Clin
Linearity Chem 10:686, 1964.
Method Direction Increase
colorimetric. Arsenazo III. Sample : Reagent Ratio 1 : 100 The reaction is linear up to calcium concentration of 20 mg/dl. 4. Peters JP, Van Slyke, DD: Quantitative clinical chemistry, vol
e.g.: Reagent volume 1 ml Specimens showing higher concentration should be diluted 1+1 2,williams and wilkins, Baltimor (MD),1932, p 760.
Assay Principle Sample volume 10 ml using physiological saline and repeat the assay (result × 2).
Temperature
o
15 - 25 C 5. Tietz NW: Blood gases and electrolytes. In:Fundamentals of
At a neutral pH, the Ca2+ form with Arsenazo III a complex, the
Zero adjustment Reagent Blank Interfering Substances: clinical chemistry, NW tietz, editor,Saunders, Philadelphia,
color intensity of which is directly proportional to the concentration of
Sensitivity 1 mg/dL (0.25 mmol/L) 176,pp 903, 908.
calcium in the sample.
Linearity 20 mg/dL (5 mmol/L) Haemolysis 6. Young DS, Effects of drugs on clinical laboratory tests. AACC
Reagents avoid haemolysis. press, Washington, D.C. 1990.
Standard Calcium (ST) Procedure
10 mg/dL 2.5 mmol/L Icterus
Blank Standard Specimen No significant interference.
Reagent (R) Standard --------- 10 ml --------- ORDERING INFORMATION
MES, pH 6.40 100 mmol/L Lipemia CATALOG NO QUANTITY
Specimen --------- --------- 10 ml No significant interference.
Arsenazo III 200 mmol/L
Reagent 1 ml 1 ml 1 ml 227 001 2 x 30 ml
Anticoagulants 227 002 2 x 100 ml
Precautions and Warnings 227 003 4 x 30 ml
o
Mix and incubate for 3 minutes at 20 - 25 C. Measure absorbance complexing Anticoagulants such as citrate, oxalate and EDTA must
be avoided.
immediately with plenty of soap and water. In case of severe injuries; of specimen (Aspecimen) and standard (Astandard) against reagent
seek medical advice immediately. blank.
Expected values
Reagent Preparation, Storage and Stability Calculation Serum, plasma
(Aspecimen)
Spectrum Calcium reagents are supplied ready-to-use and stable Serum calcium concentration (mg/dL) = x 10 Adults
up to the expiry date labeled on the bottles when stored sealed at (Astandard) 20 - 50 years 8.8-10.2 mg/dl (2.20-2.55 mmol/L)
o
15 – 25 C. (Aspecimen) >50 years 8.4- 9.7 mg/dl (2.09-2.42 mmol/L)
Urine calcium (mg/24 hrs)= x10 x10*x 2**x V***
(Astandard) Children
Deterioration 4 -18years 9.2-11.0 mg/dl (2.30-2.75 mmol/L)
Do not use the Spectrum Calcium reagents if turbid. Failure to >4 weeks 7.2-11.2 mg/dl (1.80-2.8 mmol/L)
recover control values within the assigned range may be an * The factor “10” converts mg/dl to mg/litre
indication of reagent deterioration. ** The factor “2” represents the dilution factor Urine (24 h)
*** “V” represents the 24-hour urine volume in litres Females <250 mg/day (<6.25 mmol/day)
Specimen Collection and Preservation Males <300 mg/day (<7.5 mmol/day)
Quality Control Childern <6 mg/Kg/day (<0.15 mmol/day)
Serum and plasma
Use nonhemolyzed serum.Heparin is the only acceptable antico- Normal & abnormal control serum of known concentrations should
agulant. No other anticoagulant can be used. Fresh serum collected be analyzed with each run.
in the fasting state is the perferred specimen. Serum or plasma
should be separated from cells as soon as possible, because
prolonged contact with the clot may cause lower calcium values.
Sera from patients receiving EDTA (treatment of hypercalcemia)
are unsuitable for analysis, since EDTA will chelate the calcium and
render it unavailable for reaction with O-cresolphthalein complexone.
The biological half-life of calcium in blood is few hours.
126 127
Chloride SYMBOLS IN PRODUCT LABELLING
Waste Disposal
LOT Batch Code/Lot number for use special waste collection point.
Catalogue Number Manufactured by S57: use appropriate container to avoid environmental
REF: 233 001 (4 x 25ml) 100 test REF
Consult instructions for use (T) Toxic
contamination.
REF: 233 002 (2 x 25ml) 50 test
safety data sheets.
Intended Use Urine.
Spectrum-Diagnostics Chloride reagent is intended for the in-vitro References
Urine has to be diluted 1+2 with distalled water. Multiply result
quantitative diagnostic estimation of Chloride in human serum, by 3 1. Bablok W. et al. A General Regression Procedure for Method
plasma and urine. Transformation. J Clin Chem Clin Biochem 1988;26:783-790.
System Parameters 2. Batlle DC. et al. The use of the urinary anion gap in the
Background Wavelength 492 nm (460 - 500 nm)
Chloride is the most abundant extracellular anion. Together with diagnosis of hyperchloremic metabolic acidosis. N Engl J Med
Optical path 1 cm 1988, 318:594-599.
natrium-chloride is responsible for the maintenance of osmotic Assay type colorimetric end-point
pressure, the anion-cation balance and therefore of the water Direction Increase 3. Krieg M. et al. Comparative quantitative clinico-chemical
distribution in the extracellular fluid compartment. Sample: Reagent Ratio 1:100 analysis of the characteristics of 24-hour urine and morning
Decreased plasma Cl--concentrations (hypochloremia) can result e.g.: Reagent volume 1 ml urine (in German). J Clin Chem Clin Biochem 1986, 24:863.
from salt-losing nephritis, persistent gastric secretion, prolonged Sample volume 10 ml
vomiting and metabolic acidosis that are caused by increased o o o 4. Passing H., Bablok W. A New Biometrical Procedure for
Temperature 25 C, 30 C, 37 C
production or reduced secretion of organic acids. Testing the Equality of Measurements from Two Different
Zero adjustment Against reagent blank
Increased plasma Cl--concentrations (hyperchloremia) occur Analytical Methods. J Clin Chem Clin Biochem 1983;21:709-
Linearity 130 mmol/l (462 mg/dl)
with dehydration, renal tubular acidosis, acute renal failure, in 720.
Incubation 5 min.
adrenocortical hyperfunction , salicylate intoxication and metabolic
5. Schönfeld, RG. Lewellen, CJ. A colorimetric method for
acidosis associated with prolonged diarrhoea and loss of Procedure determination of serum chloride. Clin Chem., 10, 533 (1964)
sodium bicarbonate. Chloride is often analyzed in combination
with natrium and kalium to determine the anion gap in serum and/ Pipette into clean test tubes: 6. Tietz N.W. Clinical Guide to Laboratory Tests, 3rd
or urine. The urinary anion gap is useful in the initial evaluation of Philadelphia: W.B. Saunders Company, 1995:516-519.
hyperchloremic metabolic acidosis. Blank Standard Sample
Due to the different reactivity equivalents of chloride and bromide Reagent (R) 1 ml 1 ml 1 ml
the thiocyanate method is less disturbed by the presence of bromide
than measurement with an ion- selective electrode. Standard --------- 10 ml ---------
Sample --------- --------- 10 ml ORDERING INFORMATION
Method CATALOG NO QUANTITY
Colorimetric method. Mix well, let stand for 5 minutes, then read absorbances ,Astandard
233 001 4 x 25 ml
Assay Principle and Asample against Reagent Blank at 492 nm. 233 001 2 x 25 ml
Chloride ions and Hg II-thiocyanate form thiocyanate ions in acidic
medium. Calculation
These ions react with HNO3 and Fe III-ions and effect a red colour. D Asample
The increasing extinction is directly proportional to the concentration Serum Chloride Conc.(mmol/l) = x 100
D Astandard
of chloride ions.
Expected Values
Reagents Serum 97 – 108 mmol/l.
Reagent (R) Urine 24 h urine 95 – 240 mmol/24h

Hg II - thiocyanate 2 mmol/l morning urine 54 – 158 mmol/l
Fe III - nitrate 30 mmol/l
HNO3 40 mmol/l Conversion between conventional and SI units: 1 mEq/l = 1 mmol/l
Conversion between mmol/l and mg/dl: mmol/l = 0.282 x mg/dl
Standard (S)
Chloride 100 mmol/l (354.6 mg/dl) Note:
Precautions and Warnings own reference values. The given data are only an indication.
The reagent contains mercuric thicyanate which is toxic and harmful
if inhaled or absorbed through skin. Spectrum Diagnostics does not interpret the results of a
Do not ingest or inhalate. In case of contact with eyes or skin; rinse clinical laboratory procedure; interpretation of the results is
immediately with plenty of soap and water. In case of severe injuries; considered the responsibility of qualified medical personnel.
seek medical advice immediately. All indications of clinical significance are supported by
o
Reagents and standard are ready-to-use. When stored at 2 – 8 C;
they are stable up to the expiry date stated on the label. Linearity
The assay is linear up to 130 mmol/l (462 mg/dl)
Sample
Serum.
Freshly drawn non hemolysed serum is the specimen of choice.
Chloride in serum is stable for 7 days at 2-8°C.
128 129
Carbon Dioxide (CO2) Analytical Range
The test can measure CO2 concentrations from 1.00 mmol/L up to
(Colorimetric PEPC) 50 mmol/L. At higher concentrations dilute the samples 1+1 with
NaCl solution 0.9%. Multiply result by 2.
REF: 228 001 50 Tests

228 002 100 Tests
GENERAL Waste CATALOG NO QUANTITY
Approximately 90% of total carbon dioxide present in serum is in the Handle according to the local legal regulation
form of bicarbonate. 228 001 50 Tests
Measurement of bicarbonate together with glucose, Na+, K+ and SAMPLES 228 002 100 Tests
chloride is useful in assessment of disturbances of acid base Serum, heparin plasma
balance resulting from metabolic or respiratory causes. Don’t use citrate or oxalate plasma
PRINCIPLE Stability:
Colorimetric test for the quantitative determination of Carbon Dioxide Samples should be used immediately and can be stored at 2-8°C
(CO2) in serum and plasma : four 1 hour tightly closed .
Discard contaminated samples.
Phophoenolpyruvate + Bibarbonate + NADH
Expected values
PEPC & MDH
Phosphate + Malate + NAD+ 22 – 29 mmol/L
Note: it is recommende that each laboratory should establish its own
REAGENTS reference range.
Components ( concentrations in the test ) Procedure

TRIS-Buffer (pH 7.5) Wavelength : 405 nm or 415 nm
PEP; PEPC; NADH (as reduced cofactor) Light path: 1 cm
MDH Activators, stabilizers, detergents Temperature : 37 °C
Sodium Azide 0.095% Measure decreasing absorbace against reagent blank.
Blank Sample or calibrator
Storage and stability Sample/calibrator ---- 10 µl
Reagent 1 ml 1 ml
At 2-8°C unopened reagents are stable until the expiration date
printed on the labels. After opening of bottles the stability is until Mix and incubate for 2min, read absorbance A1, and exactly after 1
the printed expiration date, if there is no contamination during the min read A2, determine ΔA = A1-A2
handling.
Calculation
D A sample
Don’t freeze reagents CO2 (mmol/L) = x Conc. Of Calibrator
D A Calibrator

Preparation of Reagents
Calibration and control
Reagent is supplied ready to use For the calibration we recommended Spectrum calibrator.
For internal QC use Spectrum control set.
Warning Take care for the sensitivity of the material against CO2 of the air.
Reagent contains Sodium azide as preservative. Don’t swallow! Calibrator concentration = 31.5 mmol/L
Don’t touch skin and/or mucous membrane
130 131
Copper SYMBOLS IN PRODUCT LABELLING o
Mix and incubate for 5 minutes at 37 C. Measure the absorbance
References
1. Abe A., Yamashita S., Noma A., Clin. Chem., 552-554-35
(Colorimetric Test EC REP Authorised Representative Temperature Limitation of the sample As and of the standard Ast against the reagent blank
(1989) 2.Richmond. N., Clin. Chem. 1973; 19: 1350-1356.
IVD For in-vitro diagnostic use Use by/Expiration Date ARBL .
with Dibrom-PAESA) LOT Batch Code/Lot number CAUTION. Consult instructions
Consult instructions for use Manufactured by DAs = As - ARBL
REF: 232 001 (2 x 25 ml ) 50 test ORDERING INFORMATION
DAstd = Astd - ARBL
REF: 232 002 ( 4 x 25 ml) 100 test CATALOG NO QUANTITY
Reagent Preparation, Storage and Stability Calculation D As 232 001 50 Test
Intended Use Spectrum Copper reagent is supplied ready-to-use and stable up to Urine Copper conc. (mg/dL)= x 5
D Astd 232 002 100 Test
Spectrum Diagnostics Copper reagent is intended for in-vitro the expiry date labeled on the bottles, Once opened, the opened vial
quantitative, diagnostic determination of Copper in human serum, o
plasma or urine on both manual and automated systems. is stable for 3 months at 2-8 C.
D As
Urine Copper conc. (mmol/l)= x 0.785
D Astd
Background Specimen collection and preparation
Copper (Cu) is an important trace element and is associated Serum, Plasma (free from haemolysis) Quality Control
with a number of metalloproteins and it is a catalytic component Normal & abnormal commercial control serum of known
of numerous enzymes and also a structural component of other 24 hours Urine: concentrations should be analyzed with each run.
important proteins. Copper is involved in many vital processes in Refrigerate or add 10 ml of 3 mol/L HCl to the conainer before
the body, Energy Production, connective tissue formation, iron collection. Linearity
metabolism, melanin synthesis, Normal function of CNS, Regulation The reaction is linear up to a Copper concentration of 500 mg/dl
of gene expression and has Antioxidant function . Excess Cu System Parameters (78.65 mmol/l)
ingestion interfere with absorbtion of zinc and can lead to Zinc Wavelength 580 nm (Hg 578) Specimens showing higher concentration should be diluted 1+1
deficiency, which is frequently characterized by slow healing, The Optical path 1 cm using physiological saline and repeat the assay (result × 2).
classical presentation of Cu toxicosis is represented by the genetic Assay type End-point
disease of Cu accumulation known as Wilson’s disease. This Direction Increase Interfering Substances:
o
disease is typified by hepatocellular damage (increase transferase) Temperature 37 C Interferences are found according to the literatures.
and/or changes in mood and behavior because of accumulation of Zero adjustment Reagent blank
Cu in Central Neurons. Linearity 500 mg/dl (78.65 mmol/l) Expected Values
In Serum
Method Procedure Adult males 70 - 140 mg/dl (11 - 22 mmol/l)
Colorimetric with Dibrom-PAESA I- Determination of copper in serum Adult females 80 - 155 mg/dl(12.5 - 24.3 mmol/l)
Blank Standard Sample Females in pregnancy 120 - 300 mg/dl (18.8 - 47 mmol/l)
Assay Principle Children (6-12 years) 80 - 190 mg/dl (12.5 - 29.8 mmol/l)
Reagent 1.0 ml 1.0 ml 1.0 ml
Copper forms with 4 - (3,5-dibromo-2-pyridylazo) - N - ethyl- Infants 20 - 70 mg/dl (3.14 - 11 mmol/l)
Standard --------- 50 ml ---------
sulfopropylaniline a chelate complex. The increase of absorbance
of this complex can be measured and is proportional to the Sample --------- --------- 50 ml In 24hours Urine 10 - 30 mg/24 hours
o
concentration of total copper in the sample. Mix and incubate for 5 minutes at 37 C. Measure the absorbance
of the sample As and of the standard Ast against the reagent blank
Reagents ARBL . considered the responsibility of qualified medical personnel.
DAs = As - ARBL All indications of clinical significance are supported by
Standard cholesterol (ST) DAstd = Astd - ARBL literature references.
100 mg/dL 15.7 mmol/L
Calculation Waste Disposal
D As
R (Monoreagent) Serum Copper conc. (mg/dL)= x 100 This product is made to be used in professional laboratories.
D Astd
Acetate buffer pH 5.0 0.2 mol/L Please consult local regulations for a correct waste disposal.
4-(3,5-dibromo-2-pyridylazo)- 0.02 mmol/L S56: dispose of this material and its container at hazardous or
D As
N-ethyl-sulfopropylaniline Serum Copper conc. (mmol/l)= x 15.7 special waste collection point.
D Astd
For further information, refer to the Copper reagent material safety contamination.
II- Determination of copper in urine
data sheet. S61: avoid release in environment. refer to special instructions/
Dilute Standard 20 Times ( Example: 50 ml standard + 950 ml normal safety data sheets.
Precautions and Warnings saline),then follow the method below :
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Blank Standard Urine Sample
Standard --------- 500 ml ---------
Avoid contamination by using clean laboratory material (pipette, Sample --------- --------- 500 ml
plastic vial for analyzers,...) Dist.H2O 500 ml --------- ---------
132 133
Total Iron SYMBOLS IN PRODUCT LABELLING
Important notes
1. Make sure that the distilled (double distilled) water is absolutely
Expected Values
Iron
Chromazurol B EC REP Authorised Representative
iron free. < 7 months : 150 – 220 mg/dL
2. Do not use turbid or hemolytic sera or plasma. Adults Women : 37 – 145 mg/dL
Single Reagent LOT
REF
Catalogue Number
for use
3. This iron test is very sensitive.To avoid contamination the Men : 55 – 175 mg/dL
glassware used must be iron free. we strongly recommend to
REF: 269 001 2 x 25 ml 50 test use disposable laboratory materials when performing this test.
REF: 269 002 4 x 25 ml 100 test 4. Bilirubin up to 15 mg/dL and copper up to 500 mg/dL do not clinical laboratory procedure; interpretation of the reults is
interfere. considered the responsibility of qualified medical personnel.
Intended Use Specimen Collection and Preservation
Performance Characteristics literature references.
Spectrum Diagnostics iron reagent is intended for the in-vitro The recommended specimen is serum or heparinized plasma.
quantitative, diagnostic determination of total iron. Plasma specimens collected with EDTA, oxalate, or citrate as Precision
anticoagulants are unsatisfactory since they bind iron, preventing Within run (Repeatiblity) Analytical Range
its reaction with the chromogen. Morning specimen is preferrable
Background Total Iron Iron : 12 – 500 mg/dl (0.9 – 89.5 mmol/ L).
to avoid low result due to diurinal variation. The biological half life of
The majority of iron in the body (~3 – 3.5 g) is found in the haemoglobin iron in blood is few hours. Level 1 Level 2
of the red blood cells or their precursors in the bone marrow. Plasma Stability: 7 days at 15 –25 oC : 3 weeks at 2 – 8 oC; n 20 20 Waste Disposal
contains very small fraction of iron (~ 2.5 mg). Iron is transported 1 year at -20 oC. This product is made to be used in professional laboratories.
Mean (mg/dL) 159 344
from one organ to another as a complex formed of ferric ions and Please consult local regulations for a correct waste disposal.
SD 2.1 1.9
a protein called apotransferrin, this iron-protein complex is called System Parameters S56: dispose of this material and its container at hazardous or
CV% 2.3 0.57
transferrin. The major iron-storage compound in the body is ferritin; it Wavelength 623 nm special waste collection point.
occurs in almost all body cells but particulary in hepatocytes. Serum Optical path 1 cm S57: use appropriate container to avoid environmental
iron is measured by the quantity of iron bound to transferrin, while Assay type End-point contamination.
Total Iron
TIBC is a direct measurement to transferrin. Elevated serum iron Direction Increase S61: avoid release in environment. refer to special instructions/
levels have been found in cases of hemochromatosis, hepatitis, Sample : Reagent Ratio 1 : 25 Level 1 Level 2 safety data sheets.
hepatic necrosis and hemolytic anemia. Decreased levels have been e.g.: Reagent volume 1 ml n 20 20
associated with iron defeciency anemia, chronic blood loss, chronic Sample volume 40 ml Mean (mg/dL) 162 351 References
disorders and insufficient dietary iron. The TIBC varies in disorders Temperature 25 oC ,30 oC or 37 oC SD 2.9 2.6 1. Bauer JD. Haemoglobin, porphyrin, and iron metabolism.
of iron metabolism so, TIBC is elevated in iron deffeciency anemia. Incubation time 5 minutes CV% 2.9 0.68 In:Kaplan LA, Pesce AJ, ed. Clinical Chemistry, theory, analysis,
The measurements of both serum iron and TIBC is fundamental in and correlation. ST. Louis:Mosby Company:1984:611-655.
evaluation and differential diagnosis of various types of anemia, liver Zero adjustment Reagent Blank Methods Comparison 2. Fairbanks VF,Klee GG. Biochemical aspects of hematology. In
disease and chronic illness. A comparison between Spectrum Diagnostics Iron reagent and a : Tietz NW, ed. Fundamentals of clinical chemistry. 3rd ed.
Sensitivity 12 mg/dL commercial reagent of the same methodology was performed on 20 Philadelphia: WB saunders:1987:789-824.
Method Linearity 500 mg/dL human sera. A correlation of 0.983 was obtained. 3. Stookey LL. Ferrozine-a new spectrophotometric reagent for
Colorimetric CAB Method. iron. Anal Chem. 1970;42:779-781.
Procedure Sensitivity 4. Viollier MA, Gschwind H, Schläpfer P. Neue
Assay Principle Reagent When run as recommended, the sensitivity of this assay is 12 mg/dL serumeisenbestimmung auf dem GSA II. Lab Med.1980;4:240-
Blank
Standard Sample 244.
Iron reacts with chromazurol B and cetyltrimethyl-ammonium bromide for serum iron.
(CTMA) to form a coloured ternary complex with an absorbance Reagent (R) 1.0 ml 1.0 ml 1.0 ml 5. Williams HL, Johnson DJ, Haut MJ. Simultaneous
measured at 623 nm.The intensity of the colour produced, is directly Linearity spectrophotometry of Fe2+ and Cu2+ in serum denatured with
Standard --------- 40 ml ---------
guanidine hydrochloride. Clin Chem.1977;23:237-240.
proportional to the concentration of iron in the sample. The reaction is linear up to iron concentration of 500 mg/dL,
Sample --------- --------- 40 ml
Specimens showing higher concentration should be diluted 1+1
Reagents Mix, and incubate for 5 minutes at 25,30 or 37 oC. Read the using physiological saline and repeat the assay (result × 2).
absorbance of the standard and sample against reagent blank. ORDERING INFORMATION
Standard Iron (ST) 200 mg/dL Interfering Substances
35.8 mmol/L CATALOG NO QUANTITY
Calculation Serum, plasma
(Asample) 269 001 2 x 25 ml
Acetate buffer PH 4.7 50 mM Iron conc. (mg/dL)= x 200 Haemolysis 269 002 4 x 25 ml
(Astandard)
CAB 0.13 mM No interference up to haemoglobin level of 5 g/L (0.3 mmol/L) in
CTMA 0.82 mM SI units determining serum iron and up to 1 g/L for TIBC.
preservatives and stabilizers ( mg/dL) x 0.1791 = mmol/L
Icterus
Precautions and Warnings Quality Control
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Normal & abnormal commercial control serum of known No significant interference up to a bilirubin level of 30 mg/dL.
immediately with plenty of soap and water. In case of severe injuries; concentrations should be analyzed with each run.
seek medical advice immediately. Lipemia
Lipemic specimens are not recommended since they may cause
Reagent Preparation, Storage and Stability negative bias. Lipemic specimens can be diluted before assay and
the dilution factor should be considered during calculation.
All reagents are are supplied ready to use and stable until expiration
date stated on label when stored refrigerated at 2 - 8 oC. Anticoagulants
Citrate, EDTA, and oxalate should be avoided.
134 135
Total Iron and Total Iron SYMBOLS IN PRODUCT LABELLING
Precision
Others
Pathological albumin levels more than 7 g/dL decrease the TIBC
Binding EC REP Authorised Representative
Within run (Repeatiblity) levels.
Expected values
Total Iron TIBC
Capacity Reagent Set LOT Batch Code/Lot number CAUTION. Consult instructions
Level 1 Level 2 Level 1 Level 2
Iron
Neonates : 36 – 184 mg/dL ( 6. 4 - 33 mmol/L)
< 7 months : 37 – 145 mg/dL (7.7 - 33 mmol/L)
Consult instructions for use Manufactured by n 20 20 20 20
Adults Women : 37 – 145 mg/dL (6.6 - 26 mmol/L)
REF: 270 001 (100 test) REF: 270 002 (100 test) Mean (mg/dL) 159 344 200 299 Men : 59 – 158 mg/dL (10.6 - 28. mmol/L)
Reagent 1 (buffer pH 4.5) SD 2.1 1.9 2.12 1.36

Reagent 1 4 x 50 ml Reagent 1 2 x 100 ml TIBC
Reagent 2 1 x 11 ml Reagent 2 1 x 11 ml Acetate buffer 0.4 mol / L CV% 2.3 0.57 1.06 0.45 1 day : 134 – 318 mg/dL (24 - 57 mmol/L)
Reagent 3 1 x 50 ml Reagent 3 1 x 50 ml Guanidine hydrochloride 1.5 mol / L 1 week : 190 – 324 mg/dL (34 - 58 mmol/L)
Reagent 4 1 x 2.5 g Reagent 4 1 x 2.5 g Hydroxylamine hydrochloride 0.6 mol / L Infants : 151 – 340 mg/dL (27 - 61 mmol/L)
Thiourea 100 mmol/L Run to run (Reproducibility) 3 – 12 months : 290 – 436 mg/dL (52 - 78 mmol/L)
Total Iron TIBC 1 – 10 years : 262 – 497 mg/dL (47 - 89 mmol/L)
Reagent 2: Ferrozine 60 mmol/L 11 – 16 years : 290 – 441 mg/dL (49 - 89 mmol/L)
Intended Use Level 1 Level 2 Level 1 Level 2
Spectrum Diagnostics iron and total iron binding capacity (TIBC) Reagent 3: Ferric chloride 120 mmol/L Adults Women : 274 – 497 mg/dL (49 - 89 mmol/L)
n 20 20 20 20
reagent set is intended for the in-vitro quantitative, diagnostic Men : 291 – 430 mg/dL (52 - 77 mmol/L)
Reagent 4: Magnesium carbonate powder. Mean (mg/dL) 162 351 203 303
determination of total iron and total iron binding capacity in human
serum. SD 2.9 2.6 2.19 1.42 Spectrum Diagnostics does not interpret the results of a
Reagent Preparation
CV% 2.9 0.68 1.12 0.51 clinical laboratory procedure; interpretation of the results is
Background Prepare the working solution by adding 5 ml of chromogen (R2) to
The majority of iron in the body (~3 – 3.5 g) is found in the haemoglobin one bottle of buffer (R1). Or prepare the working solution according
Methods Comparison All indications of clinical significance are supported by
of the red blood cells or their precursors in the bone marrow. Plasma to the number of tests required by mixing 9 volumes of R1 and
A comparison between Spectrum Diagnostics Iron and TIBC literature reference.
contains very small fraction of iron (~ 2.5 mg). Iron is transported 1 volume of R2, e.g. 900 ml R1+100 mlR2.
reagents and a commercial reagent of the same methodology was
from one organ to another as a complex formed of ferric ions and performed on 20 human sera. A correlation of 0.983 was obtained.
a protein called apotransferrin, this iron-protein complex is called Precautions and Warnings Analytical Range
transferrin. The major iron-storage compound in the body is ferritin; it Do not ingest or inhalate. In case of contact with eyes or skin; rinse Iron : 5 – 500 mg/dl (0.9 – 89.5 mmol/ L).
occurs in almost all body cells but particulary in hepatocytes. Serum immediately with plenty of soap and water. In case of severe injuries; Sensitivity TIBC : 10 – 1000 mg/dl (1.8 – 179.1 mmol/L).
seek medical advice immediately. When run as recommended, the sensitivity of this assay is 5 mg/dL
iron is measured by the quantity of iron bound to transferrin, while
for serum iron and 10 mg/dL for TIBC.
TIBC is a direct measurement to transferrin. Elevated serum iron Waste Disposal
levels have been found in cases of hemochromatosis, hepatitis, Reagent Storage and Stability This product is made to be used in professional laboratories.
hepatic necrosis and hemolytic anemia. Decreased levels have been All reagents are stable until expiration date stated on label when Linearity Please consult local regulations for a correct waste disposal.
associated with iron defeciency anemia, chronic blood loss, chronic
o
stored refrigerated at 2 - 8 C. The reaction is linear up to iron concentration of 500 mg/dL, and
disorders and insufficient dietary iron. The TIBC varies in disorders
o
Working solution is stable for 6 months at 2 – 8 C. TIBC up to concentration of 1000 mg/dl .Specimens showing higher
of iron metabolism so, TIBC is elevated in iron deffeciency anemia. concentration should be diluted 1+1 using physiological saline and
The measurements of both serum iron and TIBC is fundamental in Specimen Collection and Preservation repeat the assay (result × 2).
contamination.
evaluation and differential diagnosis of various types of anemia, liver The recommended specimen is serum or heparinized plasma. S61: avoid release in environment. refer to special instructions/
disease and chronic illness. Plasma specimens collected with EDTA, oxalate, or citrate as Procedure B- ( TIBC) safety data sheets.
anticoagulants are unsatisfactory since they bind iron, preventing 1.Pipette into a labeled tube 1 ml Reagent 3 and 0.5 ml test specimen.
o
Method its reaction with the chromogen. Morning specimen is preferrable 2.Mix thoroughly and let stand for 5–30 minutes at 15 – 25 C.
References
Guanidine / Ferrozine method. to avoid low result due to diurinal variation. The biological half life of 3.Add to each tube one spoonful of reagent 4, mix thoroughly and
o
iron in blood is few hours. leave for 30 minutes at 15 – 25 C and mix several times during the 1. Bauer JD. Haemoglobin, porphyrin, and iron metabolism.
incubation time. In:Kaplan LA, Pesce AJ, ed. Clinical Chemistry,
Assay Principle o
Stability: 7 days at 15 –25 C ; 3 weeks at 2 – 8 C;
o 4.Centrifuge at 3000 r.p.m for 10 minutes. theory, analysis, and correlation. ST. Louis:Mosby
Iron
1 year at -20 C
o 5.Carefully remove the supernatent into a clean tube and determine Company:1984:611-655.
Ferric ions are released from transferrin by guanidine hydrochloride
the serum iron as described above in procedure A within one hour.
and reduced to ferrous state by hydroxylamine. Ferrous ions 2. Fairbanks VF,Klee GG. Biochemical aspects of hematology.
react with ferrozine forming a coloured complex. To prevent copper Procedure A-( Iron ) In :Tietz NW, ed. Fundamentals of clinical chemistry. 3rd ed.
interference, cupric ions are bound to thiourea. Calculation
Reagent Standard Sample Sample Philadelphia: WB saunders:1987:789-824.
Total iron binding capacity = Iron in supernatent x 3 (dilution).
Blank Blank 3. Stookey LL. Ferrozine-a new spectrophotometric reagent for
Guanidine-HCL
Transferrin-Fe(III) Apotransferrin +Fe(III)
Working 1.0 ml 1.0 ml ------ 1.0 ml Interfering Substances iron. Anal Chem. 1970;42:779-781.
Hydroxylamine Solution Serum, plasma
Fe(III) Fe(II) 4. Viollier MA, Gschwind H, Schläpfer P. Neue
serumeisenbestimmung auf dem GSA II. Lab
Dist.water 200 ml ------ ------ ------ Haemolysis
Fe(II) + Ferrozine Colored complex Med.1980;4:240-244.
No interference up to haemoglobin level of 5 g/L (0.3 mmol/L) in
Standard ------ 200 ml ------ ------ determining serum iron and up to 1 g/L for TIBC. 5. Williams HL, Johnson DJ, Haut MJ. Simultaneous
The color intensity is directly proportional to the iron concentration spectrophotometry of Fe2+ and Cu2+ in serum denatured with
Sample ------ ------ 200 ml 200 ml
and is determined by monitoring the increase in absorbance at 546 Icterus guanidine hydrochloride. Clin Chem.1977;23:237-240.
nm. Reagent 1 ------ ------ 1.0 ml ------ No significant interference up to a bilirubin level of 30 mg/dL.
Mix, and incubate for 5 to 10 minutes at 20 – 25 C. Read the

Total Iron Binding Capacity Lipemia
The transferrin in the specimen is saturated with iron by exposure to absorbance of the standard and sample against reagent blank, and Lipemic specimens are not recommended since they may cause CATALOG NO QUANTITY
excess ferric ions; then unbound iron is removed by addition of light the absorbance of sample blank against distilled water within 30 negative bias. Lipemic specimens can be diluted before assay and
minutes at 546 nm. 270 001 4 x 50 ml
magnesium carbonate and centrifugation. The iron bound to protein the dilution factor should be considered during calculation. 270 002 2 x 100 ml
in the supernatent is measured by the Principle applied to total iron
described above. Calculation Asample - Asample blank Anticoagulants
Iron conc. (mg/dL) = x 200 Citrate, EDTA, and oxalate should be avoided.
Astandard
Reagents
Standard Iron (ST) 200 mg/dL
35.8 mmol/L Quality Control
136 137
Total Iron Binding SYMBOLS IN PRODUCT LABELLING
Serum, plasma
References
1. Bauer JD. Haemoglobin, porphyrin, and iron metabolism.
Capacity (TIBC) EC REP Authorised Representative
Use by/Expiration Date Haemolysis
In:Kaplan LA, Pesce AJ, ed. Clinical Chemistry, theory, analysis,
and correlation. ST. Louis:Mosby Company:1984:611-655.
LOT Batch Code/Lot number CAUTION. Consult instructions No interference up to haemoglobin level of 5 g/L (0.3 mmol/L) in
2. Fairbanks VF,Klee GG. Biochemical aspects of hematology. In
Catalogue Number for use determining serum iron and up to 1 g/L for TIBC.
REF : Tietz NW, ed. Fundamentals of clinical chemistry. 3rd ed.
REF: 273 001 Consult instructions for use Manufactured by Philadelphia: WB saunders:1987:789-824.
Reagent 1 1 x 50 ml
Reagent 2 1 x 2.5 g Icterus 3. Stookey LL. Ferrozine-a new spectrophotometric reagent for
No significant interference up to a bilirubin level of 30 mg/dL. iron. Anal Chem. 1970;42:779-781.
Lipemia 4. Viollier MA, Gschwind H, Schläpfer P. Neue

Procedure Lipemic specimens are not recommended since they may cause serumeisenbestimmung auf dem GSA II. Lab
Intended Use 1. Pipette into a labeled tube 1 ml Reagent 3 and 0.5 ml test Med.1980;4:240-244.
negative bias. Lipemic specimens can be diluted before assay and
Spectrum Diagnostics total iron binding capacity (TIBC) reagent is specimen. the dilution factor should be considered during calculation. 5. Williams HL, Johnson DJ, Haut MJ. Simultaneous
intended for the in-vitro quantitative, diagnostic determination of total 2.
o
Mix thoroughly and let stand for 5–30 minutes at 15 – 25 C.
iron binding capacity in human serum. spectrophotometry of Fe2+ and Cu2+ in serum denatured with
3. Add to each tube one spoonful of reagent 4, mix thoroughly Anticoagulants
o guanidine hydrochloride. Clin Chem.1977;23:237-240.
and leave for 30 minutes at 15 – 25 C and mix several times Citrate, EDTA, and oxalate should be avoided.
Background during the incubation time.
The majority of iron in the body (~3 – 3.5 g) is found in the haemoglobin 4. Centrifuge at 3000 r.p.m for 10 minutes. Others ORDERING INFORMATION
of the red blood cells or their precursors in the bone marrow. Plasma 5. Carefully remove the supernatent into a clean tube and Pathological albumin levels more than 7 g/dL decrease the TIBC CATALOG NO QUANTITY
contains very small fraction of iron (~ 2.5 mg). Iron is transported determine the serum iron as described in Total Iron Kit within levels.
from one organ to another as a complex formed of ferric ions and one hour. 273 001 50 Test
a protein called apotransferrin, this iron-protein complex is called
Expected values
transferrin. The major iron-storage compound in the body is ferritin; it Calculation TIBC
occurs in almost all body cells but particulary in hepatocytes. Serum Total iron binding capacity = Iron in supernatent x 3 (dilution). 1 day : 134 – 318 mg/dL (24 - 57 mmol/L)
iron is measured by the quantity of iron bound to transferrin, while
1 week : 190 – 324 mg/dL (34 - 58 mmol/L)
TIBC is a direct measurement to transferrin. Elevated serum iron Quality Control Infants : 151 – 340 mg/dL (27 - 61 mmol/L)
levels have been found in cases of hemochromatosis, hepatitis, Normal & abnormal commercial control serum of known 3 – 12 months : 290 – 436 mg/dL (52 - 78 mmol/L)
hepatic necrosis and hemolytic anemia. Decreased levels have been concentrations should be analyzed with each run. 1 – 10 years : 262 – 497 mg/dL (47 - 89 mmol/L)
associated with iron defeciency anemia, chronic blood loss, chronic
11 – 16 years : 290 – 441 mg/dL (49 - 89 mmol/L)
disorders and insufficient dietary iron. The TIBC varies in disorders Performance Characteristics
of iron metabolism so, TIBC is elevated in iron deffeciency anemia.
Precision Adults Women : 274 – 497 mg/dL (49 - 89 mmol/L)
The measurements of both serum iron and TIBC is fundamental in
Within run (Repeatiblity) Men : 291 – 430 mg/dL (52 - 77 mmol/L)
evaluation and differential diagnosis of various types of anemia, liver
disease and chronic illness. TIBC
Level 1 Level 2
Assay Principle clinical laboratory procedure;interpretation of the results is
The transferrin in the specimen is saturated with iron by exposure to n 20 20 considered the responsibility of qualified medical personnel.
excess ferric ions; then unbound iron is removed by addition of light Mean (mg/dL) 200 299 All indications of clinical significance are supported by
magnesium carbonate and centrifugation. The iron bound to protein literature references.
SD 2.12 1.36
in the supernatent is measured by the Principle applied to total iron
described above. CV% 1.06 0.45 Analytical Range
10 – 1000 mg/dl (1.8 – 179.1 mmol/L).
Reagents Run to run (Reproducibility)
Reagent 1: Ferric chloride 120 mmol/L Waste Disposal
TIBC This product is made to be used in professional laboratories.
Reagent 2: Magnesium carbonate powder. Please consult local regulations for a correct waste disposal.
Level 1 Level 2
Reagent Preparation, Storage and Stability n 20 20 special waste collection point.
Reagents are supplied ready-to-use and stable up to the expiry S57: use appropriate container to avoid environmental
date labeled on the bottles when properly stored refrigerated at Mean (mg/dL) 203 303
o
contamination.
2 – 8 C. SD 2.19 1.42 S61: avoid release in environment. refer to special
CV% 1.12 0.51 instructions/safety data sheets.
Do not ingest or inhalate. In case of contact with eyes or skin;
rinse immediately with plenty of soap and water. In case of severe Methods Comparison
injuries; seek medical advice immediately. A comparison between Spectrum Diagnostics TIBC reagents and a
commercial reagent of the same methodology was performed on 20
human sera. A correlation of 0.983 was obtained.
The recommended specimen is serum or heparinized plasma.
Plasma specimens collected with EDTA, oxalate, or citrate as Sensitivity
anticoagulants are unsatisfactory since they bind iron, preventing When run as recommended, the sensitivity of this assay is 10 mg/dL.
its reaction with the chromogen. Morning specimen is preferrable to
avoid low result due to diurinal variation. The biological half life of Linearity
iron in blood is few hours. The reaction is linear up to concentration of 1000 mg/dl .Specimens
showing higher concentration should be diluted 1+1 using
o
o physiological saline and repeat the assay (result × 2).
o
1 year at -20 C.
138 139
Total Iron - Ferrozine SYMBOLS IN PRODUCT LABELLING
Total Iron 1. Bauer JD. Haemoglobin, porphyrin, and iron metabolism.
REF: 272 001 (100 test) Level 1 Level 2 In:Kaplan LA, Pesce AJ, ed. Clinical Chemistry,
Reagent 1 2 x 45 ml theory, analysis, and correlation. ST. Louis:Mosby
Reagent 1B 2 x 50 ml LOT Batch Code/Lot number CAUTION. Consult instructions n 20 20 Company:1984:611-655.
Reagent 2 1 x 11 ml REF Catalogue Number for use
Mean (mg/dL) 162 351
Consult instructions for use Manufactured by 2. Fairbanks VF,Klee GG. Biochemical aspects of hematology.
SD 2.9 2.6 In : Tietz NW, ed. Fundamentals of clinical chemistry. 3rd ed.
Philadelphia: WB saunders:1987:789-824.
Intended Use Precautions and Warnings CV% 2.9 0.68
Spectrum Diagnostics iron is intended for the in-vitro quantitative, Do not ingest or inhalate. In case of contact with eyes or skin; rinse 3. Stookey LL. Ferrozine-a new spectrophotometric reagent for
diagnostic determination of total iron in human serum. immediately with plenty of soap and water. In case of severe injuries; Methods Comparison iron. Anal Chem. 1970;42:779-781.
seek medical advice immediately. A comparison between Spectrum Diagnostics Iron reagent and a
4. Viollier MA, Gschwind H, Schläpfer P. Neue
Background commercial reagent of the same methodology was performed on 20
human sera. A correlation of 0.983 was obtained. serumeisenbestimmung auf dem GSA II. Lab
The majority of iron in the body (~3 – 3.5 g) is found in the haemoglobin Reagent Storage and Stability Med.1980;4:240-244.
of the red blood cells or their precursors in the bone marrow. Plasma All reagents are stable until expiration date stated on label when
contains very small fraction of iron (~ 2.5 mg). Iron is transported o
stored refrigerated at 2 - 8 C. Sensitivity 5. Williams HL, Johnson DJ, Haut MJ. Simultaneous
from one organ to another as a complex formed of ferric ions and o
Working solution is stable for 6 months at 2 – 8 C. When run as recommended, the sensitivity of this assay is 5 mg/dL spectrophotometry of Fe2+ and Cu2+ in serum denatured with
a protein called apotransferrin, this iron-protein complex is called for serum iron.. guanidine hydrochloride. Clin Chem.1977;23:237-240.
transferrin. The major iron-storage compound in the body is ferritin; it Specimen Collection and Preservation
occurs in almost all body cells but particulary in hepatocytes. Serum The recommended specimen is serum or heparinized plasma. Linearity
iron is measured by the quantity of iron bound to transferrin, while ORDERING INFORMATION
Plasma specimens collected with EDTA, oxalate, or citrate as The reaction is linear up to iron concentration of 500 mg/dL.
TIBC is a direct measurement to transferrin. Elevated serum iron anticoagulants are unsatisfactory since they bind iron, preventing Specimens showing higher concentration should be diluted 1+1 CATALOG NO QUANTITY
levels have been found in cases of hemochromatosis, hepatitis, its reaction with the chromogen. Morning specimen is preferrable using physiological saline and repeat the assay (result × 2).
hepatic necrosis and hemolytic anemia. Decreased levels have been 272 001 100 Test
to avoid low result due to diurinal variation. The biological half life of
associated with iron defeciency anemia, chronic blood loss, chronic iron in blood is few hours. Interfering Substances
disorders and insufficient dietary iron. The TIBC varies in disorders
Serum, plasma
of iron metabolism so, TIBC is elevated in iron deffeciency anemia. o
o
The measurements of both serum iron and TIBC is fundamental in 1 year at -20 C
o
Haemolysis
evaluation and differential diagnosis of various types of anemia, liver
No interference up to haemoglobin level of 5 g/L (0.3 mmol/L) in
disease and chronic illness.
Procedure A-( Iron ) determining serum iron and up to 1 g/L .
Method Reagent Standard Sample Sample
Icterus
Guanidine / Ferrozine method. Blank Blank
No significant interference up to a bilirubin level of 30 mg/dL.
Working 1.0 ml 1.0 ml ------ 1.0 ml
Assay Principle Solution Lipemia
Iron Lipemic specimens are not recommended since they may cause
Ferric ions are released from transferrin by guanidine hydrochloride negative bias. Lipemic specimens can be diluted before assay and
and reduced to ferrous state by hydroxylamine. Ferrous ions Dist.water 200 ml ------ ------ ------
the dilution factor should be considered during calculation.
react with ferrozine forming a coloured complex. To prevent copper Standard ------ 200 ml ------ ------
interference, cupric ions are bound to thiourea. Anticoagulants
Sample ------ ------ 200 ml 200 ml
Citrate, EDTA, and oxalate should be avoided.
Guanidine-HCL Reagent 1B ------ ------ 1.0 ml ------
Transferrin-Fe(III) Apotransferrin +Fe(III)
o
Others
Hydroxylamine Mix, and incubate for 5 to 10 minutes at 20 – 25 C. Read the Pathological albumin levels more than 7 g/dL .
Fe(III) Fe(II) absorbance of the standard and sample against reagent blank, and
the absorbance of sample blank against distilled water within 30 Expected values
Fe(II) + Ferrozine Colored complex minutes at 546 nm. Iron
The color intensity is directly proportional to the iron concentration Neonates : 36 – 184 mg/dL ( 6. 4 - 33 mmol/L)
Calculation Asample - Asample blank < 7 months : 37 – 145 mg/dL (7.7 - 33 mmol/L)
and is determined by monitoring the increase in absorbance at 546 Iron conc. (mg/dL) = x 200
nm. Astandard Adults Women : 37 – 145 mg/dL (6.6 - 26 mmol/L)
Men : 59 – 158 mg/dL (10.6 - 28. mmol/L)
Reagents Quality Control Spectrum Diagnostics does not interpret the results of a
Standard Iron (ST) 200 mg/dL clinical laboratory procedure; interpretation of the results is
35.8 mmol/L considered the responsibility of qualified medical personnel.
Reagent 1 (buffer pH 4.5) All indications of clinical significance are supported by
Performance Characteristics literature reference.
Acetate buffer 0.4 mol / L
Guanidine hydrochloride 1.5 mol / L Precision
Hydroxylamine hydrochloride 0.6 mol / L Within run (Repeatiblity)
Thiourea 100 mmol/L
Analytical Range
Total Iron
5 – 500 mg/dl (0.9 – 89.5 mmol/ L).
Level 1 Level 2
Reagent 1B (buffer pH 4.5)
Acetate buffer 0.4 mol / L n 20 20
Guanidine hydrochloride 1.5 mol / L Waste Disposal
Mean (mg/dL) 159 344 This product is made to be used in professional laboratories.
Hydroxylamine hydrochloride 0.6 mol / L
Thiourea 100 mmol/L SD 2.1 1.9 Please consult local regulations for a correct waste disposal.
CV% 2.3 0.57 special waste collection point.
Reagent 2: Ferrozine 60 mmol/L
contamination.
Reagent Preparation
S61: avoid release in environment. refer to special
Prepare the working solution by adding 5 ml of chromogen (R2) to
instructions/safety data sheets.
one bottle of buffer (R1). Or prepare the working solution according
to the number of tests required by mixing 9 volumes of R1 and
1 volume of R2, e.g. 900 ml R1+100 mlR2.
140 141
MAGNESIUM SYMBOLS IN PRODUCT LABELLING Serum/Plasma:
Newborn 1.2 - 2.6 mg/dl (0.48 - 1.05 mmol/l)
Xylidyl Blue Monoreagnt EC REP Authorised Representative
Children
Women
1.5 - 2.3 mg/dl
1.9 - 2.5 mg/dl
(0.60 - 0.95 mmol/l)
(0.77 - 1.03 mmol/l)
LOT Batch Code/Lot number CAUTION. Consult instructions Men 1.8 - 2.6 mg/dl (0.73 -1.06 mmol/l)
REF: 285 001 ( 2 x 25 ml) 50 Test REF Catalogue Number for use
REF: 285 002 ( 4 x 25 ml) 100 Test Consult instructions for use Manufactured by
Urine: 1-10 mg/dl (3-5mmol/24h)
73-122 mg/24h
Specimen collection and preparation

Intended Use Serum, Plasma (free from haemolysis) and UrineThe only acceptable C.S.F.: 2.4 - 3.5 mg/dl
Spectrum Diagnostics Magnesium reagent is intended for in-vitro anticoagulant is Heparin.
quantitative, diagnostic determination of Magnesium in human Serum with any visible haemolysis cannot be used because of the
serum on both manual and automated systems. Spectrum Diagnostics does not interpret the results of a
large amount of magnesium released from the erythrocytes. The clinical laboratory procedure;interpretation of the results is
specimen should be separated from the clot as soon as possible to considered the responsibility of qualified medical personnel.
Background prevent falsely elevated magnesium due to passage of magnesium All indications of clinical significance are supported by
Magnesium is an activator for various physiochemical processes, from the erythrocytes into the serum. literature references.
including phosphorylation, protein synthesis, and DNA metabolism. EDTA, Sodium fluoride and oxalate should be avoided because they
It is also involved in neuromuscular conduction and excitability of interfere with the results.
skeletal and cardiac muscle. Dynamic Range
Ingested magnesium is absorbed in the intestine and the amount System Parameters 0.2 - 5.0 mg/dl.
absorbed is inversely related to the total magnesium intake. T he Wavelength 546 nm (500 – 550 nm)
kidneys effectively control magnesium homeostasis through tubular Optical path 1 cm Waste Disposal
reabsorption, which conserves magnesium when intake is low and Assay type End-point This product is made to be used in professional laboratories.
excretes excess when intake is high. Direction Increase Please consult local regulations for a correct waste disposal.
Increased serum magnesium concentrations occur in renal failure, Temperature 25 C
o
acute diabetic acidosis, dehydration, or Addison’s disease. Zero adjustment Reagent blank special waste collection point.
Hypermagnesemia has a depressing effect on the central nervous Sensitivity 0.2 mg/dL S57: use appropriate container to avoid environmental
system, causing general anesthesia and respiratory failure. It alters Linearity 5.0 mg/dL contamination.
the conduction mechanism of the heart, causing cardiac arrest.
Hypomagnesiumia may be observed in chronic alcoholism,
Procedure safety data sheets.
malabsorption, severe diarrhea, acute pancreatitis, diuretic
therapy, prolonged parenteral fluid therapy without magnesium Blank Standard Sample
supplementation, and the kidney disorders such as glomerulonephritis References
and tubular reabsorption defects.
Standard ------ 10 µl ------ 1. Thomas L. Clinical Laboratory Diagnostics 1st ed Frankfurt:
Method TH-Books Verlagsgesellschaft; 1998. p. 231-41.
Sample ------ ------ 10 µl
Xylidyl Blue, Colorimetric Endpoint. 2. Mann ck,yoe JH. Spectrophotometric determination of Mg
Mix well and let stand 10 minutes at room temperature , then read Anal.chem Acta 1957; 16: 155 - 60
Assay Principle absorbance of sample and standard against reagent blank.
Magnesium ions form a colored chelate complex when reacting The color is stable for at least 1 hour.
with xylidyl blue in alkaline solution, the intensity of the color is
proportional to the magnesium concentration. Calcium ions are ORDERING INFORMATION
Calculation
masked by GEDTA. CATALOG NO QUANTITY
(Aspecimen)
Serum Magnesium conc. (mg/dL)= x 2.5 285 001 2 x 25 ml
Reagents (Astandard)
285 002 4 x 25 ml
Standard cholesterol (ST) Quality Control
2.5 mg/dL 1.0 mmol/L ( 1.03mmol/l ) Normal & abnormal commercial control serum of known
R: Reagent
Methods Comparison
Ethanolamine pH 11.0 1 mol/L A comparison between Spectrum Diagnostics Magnesium reagent
GEDTA (Glycoletherdiamin-tetraacetic acid) 60 mmol/L and a commercial reagent of the same methodology was performed
Xylidylblue 110 mmol/L on 20 human sera. A correlation of 0.995 was obtained.

For further information, refer to the Magnesium reagent material
Sensitivity
safety data sheet.
assay is 0.2 mg/dL.
Linearity
The reaction is linear up to a Magnesium concentration of 5.0 mg/
dl; specimens showing higher concentration should be diluted 1+1
using physiological saline and repeat the assay (result × 2).
Avoid contamination by using clean laboratory material (pipette,
plastic vial for analyzers,...)
Interfering Substances:
Interferences are found according to the literature.
The reagents are supplied ready to use.
Magnesium reagent is stable up to the expiry date labeled on the Expected Values
o
bottles when stored at 2 - 8 C. The following guidelines may be used for clinical interpretation:
142 143
Phosphorus, Inorganic SYMBOLS IN PRODUCT LABELLING
Quality Control
Analytical Range
1 – 20 mg/dL.
EC REP Authorised Representative Use by/Expiration Date concentrations should be analyzed with each run.
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions Waste Disposal
REF: 294 001 (4 x 25 ml) 100 test
LOT Batch Code/Lot number for use Performance Characteristics This product is made to be used in professional laboratories. Please
REF: 294 002 (2 x 100 ml) 200 test
Catalogue Number Manufactured by Precision consult local regulations for a correct waste disposal.
REF: 294 003 (4 x 100 ml) 400 test REF
Within run (Repeatiblity) special waste collection point.
Level 1 Level 2 contamination.
Intended Use Specimen Collection and Preservation S61: avoid release in environment. refer to special instructions/
Spectrum Diagnostics phosphorus reagent is intended for the in- n 20 20
safety data sheets.
vitro quantitative, diagnostic determination of phosphorus in human Serum and plasma Mean (mg/dL) 5.49 8.92
serum and urine on both automated and manual systems. Serum specimens collected from fasting patients are the preferable SD 0.21 0.128
References
specimens since meals lower inorganic phosphate level. The 1. Daly JA, Ertingshausen G: Direct method for determination
Background CV% 3.83 1.43
heparin is the only acceptable anticoagulant. EDTA, fluoride and of inorganic phosphate in serum with the centerifichem. Clin
The body of human adult contains approximately 620 g of phosphorus citrate lower the inorganic phosphate level.Serum and plasma Chem 18:263, 1972.
entirely in the form of phosphates distributed fairly equally between should be separated from erythrocytes as soon as possible to avoid
extracellular and intracellular compartments. About 85 % of Run to run (Reproducibility) 2. Frankel S:Electrolytes. In: Gradwhol’s clinical laboratory
the leakage of inorganic phosphate and phosphate esters into the
extracellular phosphate occurs in inorganic forms as hydroxyapatite. plasma media. The biological half-life in serum is few minutes. Level 1 Level 2 methods and diagnosis, 6 th ed. S Frankel, S Reitman,
In plasma or serum, most phosphate exist in the inorganic form o
Stability: 1 day at 15 – 25 C ; 4 days at 4 – 8 C;
o
Editors, Mosby, St. louis (MO), 1963, p 188, 1963 .
(Pi); this fraction is present as the mono-and dihydrogen forms, the o n 20 20
1 year at -20 C
relative proportions varying with the pH. Intracellularly, phosphate 3. Hanok A, Kao J: The stability of a reconstituted serum for the
Urine Mean (mg/dL) 5.61 9.1
occurs mainly as phospholipids and phosphoproteins; this fraction is assay of fifteen chemical constituents.Clin Chem 14:58, 1968 .
Urine specimens should be collected in an acid-washed, detergent- SD 0.29 0.133
termed organic phosphate. Increased serum phosphate levels may free container. Acidify with hydrochloric acid after collection (pH<3). 4. young DS: Effects of drugs on clinical laboratory tests. 3 ed
occur in renal failure, hypervitaminosis D,and hypoparathyroidism. o
Stability : 2 days at 15 – 25 C ; 6 months at 2 – 8 C;
o CV% 3.97 1.5 ed., AACC press, Washington (DC), 1990; Supplement No. 1,
Reduced serum phosphate levels are seen in vtamin D deficiency, Urine samples should be diluted 1 : 10 ( 1 + 9 ) with distilled water 1991 .
rickets, hyperparathyroidism, and fanconi’s syndrome. before assay ; multiply the result by 10.
Methods Comparison
Method System Parameters A comparison between Spectrum Diagnostics Phosphorus reagent
UV – phosphomolybdate method. and a commercial reagent of the same methodology was performed ORDERING INFORMATION
Wavelength 340 nm
Optical path 1 cm on 20 human sera. A correlation of 0.947 was obtained. CATALOG NO QUANTITY
Assay principle Assay type End-point
294 001 4 x 25 ml
Inorganic phosphate reacts with ammonium molybdate in presence Direction Increase Sensitivity
294 002 2 x 100 ml
of sulfuric acid to form nonreduced phosphomolybdate. Sample : Reagent Ratio 1 : 100 When run as recommended, the minimum detection limit of this
294 003 4 x 100 ml
e.g.: Reagent volume 1 ml assay is 1 mg/dL.
H2SO4 Sample volume 10 ml
Phosphate + Ammonium molybdate Nonreduced
phosphomolybdate Temperature
o
15 – 25 C or 37 C
o
Linearity
Zero adjustment Reagent blank The reaction is linear up to phosphorus concentration of 20 mg/dL.
o
the concentration of phoshomolybdate formed is directly proportional Incubation time 10 minutes at 15 – 25 C or Specimens showing higher concentration should be diluted 1+4
o
to the inorganic phosphate concentration. It is determined by 5 minutes at 37 C using physiological saline and repeat the assay (result × 5).
measuring the increase in absorbance at 340 nm. Reagent Blank Limits Low 0.00 AU
High 0.5 AU Interfering Substances
Reagents Sensitivity 1 mg/dL (Serum, plasma)
Standard phosphorus (ST) Haemolysis
5 mg/dl 1.61 mmol/L Procedure Avoid haemolysis since RBCs contain very high levels of inorganic
Reagent (R) Blank Standard Sample phosphate .
ammonium molybdate 3.5 mmol/L
Reagent 1 ml 1 ml 1 ml Icterus
sulphuric acid 750 mmol/L
Surfactants 1% Distilled water 10 µl ------ ------ No significant interference up to a bilirubin level of 30 mg/dL.
(C)-Corrosive contains caustic materials. Standard ------ 10 µl ------
R35 Causes severe burns. Lipemia
R41 Risk of serious damage to eyes. Sample ------ ------ 10 µl No significant interference.
S26 In case of contact with eyes, rinse immediately with
o
Mix, wait for 10 minutes at 15 – 25 C or 5 minutes at 37 C, then
o
Anticoagulants
EDTA, citrate and fluoride interfere with the test .
S28 After contact with skin, wash immediately with plenty of measure absorbance of specimen (Aspecimen) and standard
soap and water.
(Astandard) against reagent blank within 30 minutes. Expected Values
For further information, refer to the Phosphorus, Inorganic reagent Serum (fasting)
material safety data sheet. Calculation (Aspecimen)
Serum phosphorus conc. (mg/dl) = x 5 Adults : 2.7 – 4.5 mg/dL (0.87 –1.45 mmol/L)
(Astandard) Children< 12 years : 4.5 – 5.5 mg/dL (1.45 – 1.78 mmol/L)
Precautions and Warnings Children < 1year : 4.5 – 6.7 mg/dL (1.45 – 2.16 mmol/L)
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Neonates : 5.0 – 9.6 mg/dL (1.60 – 3.10 mmol/L)
immediately with plenty of soap and water. In case of severe injuries; (Aspecimen) Urine ( 24 hrs) : 0.3 – 1.0 g/24 hrs (11 – 32 mmol / day)
Urine phosphorus conc. (mg/dl) = x 5 x 10
seek medical advice immediately. (Astandard)
Reagent Storage and Stability Note: For turbid highly icteric sera , prepare a seum blank by adding clinical laboratory procedure;interpretition of the results is
Reagents are supplied ready-to-use and stable until expiration date
o 10 ml serum to 1 ml saline into a labeled test tube. Read absorbance considered the responsibilaty of qualified medical personnel.
stated on label when stored refrigerated at 2 - 8 C
of serum blank at 340 nm vs water and subtract from test absorbance All indications of clinical significance are supported by
before calculating results. literature references.
Deterioration
If absorbance of reagent measured at 340 nm against distilled water
as reference is greater than 0.5, it should be discarded.
144 145
INORGNIC PHOSPHORUS Blank Standard Sample
Lypemia
No significant interference
R1 0.5 ml 0.5 ml 0.5 50ml CATALOG NO QUANTITY
REF 296 001 2x 50 ml 100 test Drugs
REF 296 002 2x100 ml 200 test R2 0.5 ml 0.5 ml 0.5 ml 296 001 2x 50 ml
Young in 1990 has published a comprehensive list of drugs and 296 002 2x 100 ml
Standard (µL) ------ 50 µL ------ substances which may interfere with this assay.
Colorimetric method without deproteinization
Sample (µL) ------ ------ 50 µL
Warning & Precaution
Mix and Incubate for 15 min at 20-25°C. Then Read the absorbance SPECTRUM INORGANIC PHOSPHORUS reagent is for in-vitro
INTRODUCTION & PRINCIPLE diagnostics use only. Normal precautions with handling laboratory
of the sample (A1) and Standard (A2) against the Reagent Blank.
Spectrum Inorganic Phosphorus Reagent Used For The In-Vitro reagents should be followed.
(The color is stable for 60 mins.
Quantitative Determination Of Inorganic Phosphorus In Human Reagent is an acid and is caustic, so, avoid contact with skin. Flush
Serum, Plasma And Urine On Both Of Manual And Automation. with plenty of water if contact occurs.
The Measurement Of Inorganic Phosphorus In Serum Is Usually CALCULATION
Don´t pipette by mouth.
Accomplished By Forming A Phosphomolybdate Complex And In It is recommended that disposable the PVC tubes are used for this
Turn Reducing It To A Molybdenum blue Color Complex Methods A1 Sample
X Standard conc. = mg/dl of inorganic phosphorus assay.
Differ As To Choice Of Reducing Agent Stannous chloride,
Phenyl hydrazine, Aminonaphthoisulfonic Acid, Ascorbic Acid, A2 Standard
Imprecision
Pmethylaminophenolsulfte, N-Phenyl-P-Phenylenediamine And
Unit Conversion Reproducibility was determined using in an internal protocol.
Ferrous Sulfate. These Methods Suffered From Color Instability.
mmol/l =mg/dl x 0.323 The following results were obtained
Deproteinization Steps And Complexity Of Performance. Spectrum
Regents Eliminated The Need To Prepare A Protein Free Filtrate,
Accelerated Color Production, Stabilized The Color And Simplified CALCULATION for 24 hr Urine Within Run
The Procedure: Inorganic Phosphorus Reacts With Ammonium
Control Level 1 Level 2
Molybdate In An Acidic Medium To Form A Phosphomolybdate A1 Sample
Complex, Whic Reduced By Ferrous Ammonium Sulfte To Produce X 5 x10 x V = mg/day of inorganic phosphorus No. of Samples 50 50
A Molybdenum Blue Complex. A2 Standard
Mean(mg/dl) 4.0 6.8
The Intensity Of Color Measured Photometrically At 600 – 675
Nm, And Its 00000000000 Is Directly Proportional To Inorganic Where: SD(mg/dl) 0.03 0.06
Phosphorus Concentration In The Specimen. 5.0 Standard concentration CV (%) 2.7 2.7
10 Dilution Factor
REAGENTS V 24 hr.urine volume in litre
Between Day
S PHOSPHORUS STANDARD 5.0mg/Dl
R1 Reducing Reagent Expected Values Control Level 1 Level 2
SULFURIC ACID 1.1 N
No. of Samples 50 50
FERROUS AMMONIUM SULFATE 100 G/L For Fasting Serum
FERROUS NITRATE 2.0 G/L Mean(mg/dl) 4.2 6.0
Children <2 years 4.5-6.7 mg/dl (1.45-2.16 mmol/l)
R2 Color Reagent SD(mg/dl) 0.07 0.09
AMMONIUM MOLYBDATE 4.5 G/L Children 2-12 years 4.5-5.5 mg/dl (1.45-1.78 mmol/l)
SULFURIC ACID 1.1 N CV (%) 4.9 2.6
Adults >12 years 2.5-5.0 mg/dl (0.81-1.62 mmol/l)
REAGENTS PREPARATION AND STABILITY For Urine

All reagents are supplied ready to use and stable up to the expiry Method Comparison
o On Non restricted diet 0.4-1.3 g/day (12.9-42 mmol/l) Comparison studies were carried out by using similar commercially
date given on the label When stored at 2-8 C
available inorganic phosphorus reagents as a reference.
Linearity Ser m, plasma (Heparin) and urine samples were assayed in parallel
Specimen Collection and preservation
The assay is linear up to 14 mg/dl Above this value should be diluted and the results compared by least squares regression.
SPECTRUM Inorganic phosphorus kit can be used with
with 0.9% NaCl Or Distilled water and re-assayed, then multiply the The following statistics were obtained:
Plasma, serum and urine.
Result by the dilution factor.
Serum and Plasma SERUM/PLASMA
Fresh serum collected in the fasting state is the preferred Sensitivity No. of sample pairs 50
specimen, since serum inorganic phosphorus level are lower The sensitivity of the inorganic phosphorus Is 0.3 mg/dl Range of sample results 2.5- 7.4 mg/dl
after meals. Heparin is the only anticoagulant which can be used Mean of reference method results 4.3 mg/dl
and serum or plasma should be separated from blood cells as Quality Control Mean of inorganic phosphorus results 4.1mg/dl
soon as possible to avoid the leakage of inorganic phosphorus and It is recommended that controls (normal and abnormal) be included Slope 0.93
phosphate esters into the plasma media. in each set of assays, or at least once a start, or, when a new reagent Intercept o.21mg/dl
o
Inorganic phosphorus in serum is stable for 7 days at 4 C and is used, or, after preventive maintenance is performed or a clinical Correlation Coefficient 1.00
For 3 weeks when frozen. component is replaced.
Interfering Substances BIBLIOGRAPHY

Urine
Urine specimens should be collected in acid-washed containers 1. Fraser et al., (1987) calcium and phosphate metabolism
(5ml of 6.0mol/l HCL). Anticoagulants in Tietz, NW.ed. Fundamental ofclinicalchemistry.3₫ed.
Stored Urine specimen must be mixed well and diluted 1:10 in The only accepted one is the heparin. Philadelphia:705-728
distilled water prior analysis and Inorganic phosphorus stable if Complexing anticoagulants such as EDTA, CITRATE AND OXALATE
stored at room temp., refrigerated or frozen. Must be avoided. 2. Tietz, NW. ed.,(1990) Clinical Guide to Laboratory Tests,2nd
Philadelphia Pa: WB Saunders company:444-447
PROCEDURE BILIRUBIN 3. Young, DS (1990): Effects of Drugs on. Clinical Laboratory
Wavelength 600-675 nm No significant interference from free or conjugated bilirubin up to a Tests Third Edition 1990:3:6-12.
Cuvette 1 cm light path level 30mg/dl.
o
Temperature 20-25 C
Zero adjustment Reagent Blank Haemolysis
Specimen Serum, Plasma or Avoid haemolysed specimen since high amount of phosphate is
Diluted Urine liberated during rupture of erythrocytes.
146 147
Potassium SYMBOLS IN PRODUCT LABELLING
Waste Disposal
Tetraphenylborate Method EC REP Authorised Representative
Without Deproteinization LOT
REF
Catalogue Number
for use
Manufactured by

S57:
contamination.
REF: 298 001 (2 x 25ml) 50 test
REF: 298 002 (4 x 25ml) 100 test Temperature Limitation
safety data sheets.
REF: 298 003 (2 x 100ml) 200 test
References
Intended Use System Parameters
Spectrum-Diagnostics Potassium reagent is intended for the in-vitro Wavelength 578 nm 1. Hillman, G.; Beyer, G.: Z. Klin. Biochem. 5 (1967), 93
quantitative diagnostic estimation of potassium in human serum on Optical path 1 cm
Assay type Colorimetric end-point 2. Hoeflmayr, J.: Praxis und Helferin 8 (1979)
manual systems.
Direction Increase 3. Tietz, N.W.: Fundamentals of Clin. Chem. (1976), 876
Background
Sodium and Potassium are the major cations of extracellular and
Sample volume 20 ml
intracellular fluids respectively. Sodium maintains the normal o
Temperature 25-37 C
distribution of water and the osmotic pressure in the various fluid
Equilibration Time 30 seconds CATALOG NO QUANTITY
compartments. Potassium influences the acid base balance and
Zero adjustment Against reagent blank
osmotic pressure including water retention. Increased sodium 298 001 2 x 25 ml 50 Test
Linearity 10 mmol/L
levels are found in severe dehydration and excessive treatment 298 002 4 x 25 ml 100 Test
with sodium salts. Decreased levels are found in severe polyurea, 298 003 2 x 100 ml 200 Test
metabolic acidosis, diarrhea and renal insufficiency. Increased Procedure
potassium levels are found in renal failure, dehydration, shock and Reagent Blank Standard Sample
adrenal insufficiency. Decreased levels are found in malnutrition,
Reagent R 1mL 1mL 1mL
gastrointestinal fluid loss, and hyperactivity of the adrenal cortex.
Standard ------ 20 µl ------
Method Sample ------ ------ 20 µl
Turbidimetric Tetraphenylborate (TPB)
o o
Mix, incubate for 3 minutes at 37 C or 5 minutes at 25 C,
Assay Principle
Mix again thoroughly and read absorbance of sample (Asample) and
At an alkaline pH Potassium ions and TPB form a turbid emulsion, the
increase of which can be measured quantitatively in a photometer at standard (Astandard) against blank.
578 nm. The increase of the absorbance (A) is directly proportional
to the concentration of Potassium in the sample. Calculation
(Asample)
Reagent Serum Potassium Conc.(mmol/L) = x5
A
( standard)
Reagent R NaOH 0.50 mol/L Expected Values
TPB-Na 240 mmol/L Serum 3.6 - 5.5 mmol/L
Plasma 4.0 – 4.8 mmol/L
Irritant (Xi): R36/38: Irritating to eyes and skin. S26: In case of
contact with eyes, rinse immediately with plenty of water and Note:
seek medical advice. S37/39: Wear suitable gloves and eye/face It is recommended for each laboratory to establish and maintain its
protection. own reference values. The given data are only an indication.
Standard Potassium 5.00 mmol/L

Precautions and Warnings considered the responsibility of qualified medical personnel.
Do not ingest or inhalate. In case of contact with eyes or skin; rinse All indications of clinical significance are supported by
immediately with plenty of soap and water. In case of severe injuries; literature references.
Reagent Storage and Stability Linearity

o
Reagent and standard are ready-to-use. When stored at 2 – 25 C; The assay is linear up to 10 mmol/L
they are stable up to the expiry date printed on the label. The
remaining stability after opening the bottles is 1 month at Interfering substances
o
18 – 25 C Interferences are found according to the literature.
Samples
Human Serum.
148 149
Sodium SYMBOLS IN PRODUCT LABELLING
Linearity
The assay is linear up to 180 mEq/l.
LOT Batch Code/Lot number CAUTION. Consult instructions Waste Disposal

REF: 303 001 (2 x 25ml) 50 test REF Catalogue Number for use This product is made to be used in professional laboratories.
REF: 303 002 (4 x 25ml) 100 test Consult instructions for use Manufactured by Please consult local regulations for a correct waste disposal.
REF: 303 003 (2 x 100ml) 200 test S56: dispose of this material and its container at hazardous or
contamination.
Intended Use Serum or heparinised plasma, CSF & Urine. Urine diluted 1+1with
Spectrum-Diagnostics Sodium reagent is intended for the in-vitro distilled water can be used for Chloride estimation. Chloride in
safety data sheets.
quantitative diagnostic estimation of sodium in human serum on serum is stable for 7 days at 2-8 °C.
manual systems. References
Background System Parameters 1. Tietz, N.W., Fundamentals of clinical Chemistry, W.b. Saunders
Sodium and Potassium are the major cations of extracellular and Wavelength 630 nm Co. Phila, P.A. p. 874.
intracellular fluids respectively. Sodium maintains the normal Optical path 1 cm 2. Henry R.F., et, al, Clinical Chemistry Principles and Technics.
distribution of water and the osmotic pressure in the various fluid Assay type colorimetric end-point 2nd Ed, Harper and Row, Harper and Row, Hargersein, M.D.
compartments. Potassium influences the acid base balance and Direction Increase (1974)
osmotic pressure including water retention. Increased sodium Sample: Reagent Ratio 1:100
levels are found in severe dehydration and excessive treatment e.g.: Reagent volume 1 ml 3. Maruna RFL., Clin Chem. Acta. 2:581, (1958)
with sodium salts. Decreased levels are found in severe polyurea, Sample volume 10 ml 4. Trinder, P:Analyst, 76:596, (1951)
metabolic acidosis, diarrhoea and renal insufficiency. Increased Temperature Room temperature
potassium levels are found in renal failure, dehydration, shock and Zero adjustment Against reagent blank
adrenal insufficiency. Decreased levels are found in malnutrition, Linearity 180 mEq/l
gastrointestinal fluid loss, and hyperactivity of the adrenal cortex. Incubation 5 min. ORDERING INFORMATION
Blank absorbance limit 1.2
CATALOG NO QUANTITY
Method
Colorimetric method. Procedure 303 001 2 x 25 ml 50 Test
303 002 4 x 25 ml 100 Test
Assay Principle Pipette into clean test tubes: 303 003 2 x 100 ml 200 Test
The Present method is based on reaction of sodium with a selective
chromogen producing a chromophore whose absorbance varies
directly as the concentration of sodium in the test specimen Reagent (R) 1mL 1mL 1mL
Standard ------ 10 µl ------
Reagents
Sample ------ ------ 10 µl
Reagent (R) Color Reagent
Mix well, let stand for 5 minutes at R.T.
Chromogen 0.03 gm/L then read absorbances ,Astandard and Asample against
EDTA 25 mmol/L Reagent Blank at 630 nm.
Bicine buffer 75 mmol/L
preservatives 0.05% Calculation
Antifoam 0.01 %
(Asample)
Serum Sodium Conc.(mEq/l) = x 150
Standard (S) Sodium 150 mEq/l (Astandard)
Precautions and Warnings Expected Values

Do not ingest or inhalate. In case of contact with eyes or skin; rinse Serum 135 – 150 mEq/l.
seek medical advice immediately. Note:
Reagent Storage and Stability own reference values. The given data are only an indication.
Reagents and standard are ready-to-use.
o
When stored at 2 – 25 C; they are stable up to the expiry date stated
on the label. The remaining stability after opening the bottles is
o Spectrum Diagnostics does not interpret the results of a
1 month at 2 – 25 C.
Sample All indications of clinical significance are supported by
Freshly drawn non hemolysed serum is the specimen of choice. literature references.
Serum Sodium are is stable for at least 24 hours at room temperature
and two weeks at 2-8°C.
150 151
ZINC SYMBOLS IN PRODUCT LABELLING
Waste Disposal
(Colorimetric Test EC REP Authorised Representative
with 5-Brom-PAPS) LOT
REF
Catalogue Number
for use

S57:
contamination.
REF: 330 001 ( 2 x 25 ml) 50 test
safety data sheets.
REF: 330 002 ( 4 x 25 ml) 100 test
System Parameters Spectrum Diagnostics does not interpret the results of a

Intended Use Wavelength 560 nm clinical laboratory procedure; interpretation of the results is
Spectrum Diagnostics liquizyme Zinc reagent is intended for the in- considered the responsibility of qualified medical personnel.
Optical path 1 cm
vitro quantitative, diagnostic determination of Zinc in human serum, All indications of clinical significance are supported by
Assay type Colorimetric End point
plasma or Urine. literature references.
Background Sample volume 50 ml
Zinc is an essential element in the nutrition of human beings, zinc Temperature 25 C or 37 C
o o
is required in the genetic make-up of every cell and is an absolute Zero adjustment Against Reagent Blank (RBL) References
requirement for all biologic reproduction. Linearity 400 mg/dl (61.2 mmol/l)
Zinc is needed in all DNA and RNA syntheses and is required at 1. Johnsen and R.Eliasson. Evaluation of a commercially available
every step of the cell cycle. kit for the colorimetric determination of zinc.International
Procedure Journal of Andrology, 1987, April 10 (2):435-440.
About 2 grams of zinc is distributed throughout the body human.
2. Tietz, text book of clinical chemistry and molecular diagnostics
Hypozincemia is a condition where insufficient zinc is available for Reagent (R) 1mL 1mL 1mL ISBN 0-7216-0189-8
metabolic needs. The defficiency may lead to Anorexia, Diarrhia and
Pneumonia or cognitive and motor function impairment in children. Standard (St) ------ 50 µl ------
Sample ------ ------ 50 µl
Zinc defficiency during pregnancy can negatively affect both the
mother and fetus. o
Mix and incubate for 10 min at 25 C or 5 min at 37 C.
o
Measure the absorbance of the sample As and the absorbance of
Method standard Ast against reagent blank. CATALOG NO QUANTITY
Colorimetric Method with 5-Brom-PAPS.
330 001 2 x 25 ml
Calculation 330 002 4 x 25 ml
Assay Principle Aspecimen
Zinc forms with 2-(5-Brom-2-pyridylazo)-5-(N-propyl-N- Zinc Concentration (mg/dl) = x 200
sulfopropylamino)-phenol a red chelate complex. Astandard
The increase of absorbance can be measured and is proportional
to the concentration of total zinc in the sample. Aspecimen
Zinc Concentration (mmol/l) = x 30.6
Astandard
Reagents
Standard (St.) 200 mg/dl (30.6 mmol/l) Quality Control

Reagent (R) be analyzed with each run.
5-Br-PAPS 0.02 mmol/L
Bicarbonate buffer pH 9.8 200 mmol/L Methods Comparison
Sodium Citrate 170 mmol/L A comparison between Spectrum Diagnostics Zinc reagent and a
Dimethylglyoxime 4 mmol/L commercial reagent of the same methodology was performed on 20
Detergent 1% human serum.A correlation of 0.993 was obtained.
For further information, refer to the Aspartate aminotransferase Linearity

reagent material safety data sheet. The reaction is linear up to Zinc concentration of 400 mg/dl.
Precautions and Warnings Expected values

Do not ingest or inhalate. In case of contact with eyes or skin; rinse Serum/Plasma
seek medical advice immediately. Male: 72.6 - 127 mg/dl (11.1 - 19.5 mmol/l)
Female: 70.6 - 114 mg/dl (10.7 - 17.5 mmol/l)
Reagent Preparation During pregnancy and menstruation the concentration of zinc can
Spectrum Zinc reagents are supplied ready-to-use. be very low
Children: 63.8 - 110 mg/dl (9.8 - 16.8 mmol/l)
Reagent Storage and Stability New born: 49.5 - 99.7 mg/dl (7.6 - 15.3 mmol/l)
o
stored refrigerated at 2 - 8 C. Urine
300 - 800 mg/dl
Specimen
Serum, Plasma or Urine
152 153
B-Quality control
154 155
SP-NORMOTROL SYMBOLS IN PRODUCT LABELLING ORDERING INFORMATION
Control serum normal/borderline, for Control EC REP Authorised Representative Temperature Limitation CATALOG NO QUANTITY
of Accuracy and Precision in quantitative IVD For in-vitro diagnostic use Use by/Expiration Date 326 001 (1 vial ) 1 x 5 ml
LOT Batch Code/Lot number CAUTION. Consult instructions 326 002 (5 vials) 5 x 5 ml
In vitro Diagnostics REF Catalogue Number for use
REF: 326 001 (1 vial ) 1 x 5 ml

REF: 326 002 (5 vials) 5 x 5 ml
Description Expected Values

SP-NORMOTROL is a lyophilized human-based control serum.
1. The expected value of specific assays are provided on the
Storage and stability assay value sheet accompanying each kit, and are lot specific.
Unopened bottles must be stored at 2 - 8°C and are stable up to
the expiration date printed on the labels 2. The expected values are obtained using replicate assay of
each manufactured lot of SP-Normotrol.
After reconstitution the serum can be used , stored at 2 - 8°C and
protected from light , in between 5 days. 3. The individual laboratory values should fall within the expected
Bilirubin is stable 1 day ,refrigerated . values.
The frozen serum is stable for at least 30 days at < -20°C .
4. It must however be noted that each laboratory should establish
Warnings and Precautions its own normal values and reference range according to GLP.
Each individual blood donation used for this serum was found
negative when tested with FDA-approved methods on HBsAg, anti- Literature
HIV 1+2 and anti-HCV. Nevertheless the serum should be treated
for safety reasons always as a potentially infectious material . 1. Röhle G, Siekmann L. Quality assurance of quantitative
determination. In: Thomas L, editor. Clinical laboratory
Preparation diagnostics.1st ed. Frankfurt:TH-Books Verlagsgesellschaft;
Reconstitute carefully by adding exactly 5 ml of distilled water. 1998. p. 1393-1401
Allow the vial to stand for 30 minutes ,swirling occasionally.
Avoid foaming i.e. do not shake! 2. Biosafety in Microbiological and Biomedical Laboratories. U.S.
For alkaline phosphatase the control serum should stand for Department of Health and Human Services, Washington 1993
2 hours at +25 °C before use . (HHS Publication No. [CDC] 93-8395).
Procedure 3. Richtlinien der Bundesärztekammer zur Qualitätssicherung in

Refer to the package inserts of the reagent kits medizinischen Laboratorien. Deutsches Ärzteblatt 1988;85:
B519–B532.
Assay Values and Ranges
Assay values for analytes for which approved reference methods 4. Richtlinie der Bundesärztekammer zur Qualitätssicherung
are available were determined according to Guidelines of the quantitativer laboratoriumsmedizinischer Untersuchungen
German Federal Medical Council [Bundesaerztekammer] from 1987 Deutsches Ärzteblatt 2002;98:A 2747-59.
(reference method values) [3].
Ranges of acceptance were calculated as assigned value ± three
times the maximum tolerable deviation of a single value according to
the Guidelines of the German Federal Medical Council [4].
156 157
SP-PATHOTROL SYMBOLS IN PRODUCT LABELLING ORDERING INFORMATION
Control serum pathological, for Control EC REP Authorised Representative Temperature Limitation CATALOG NO QUANTITY
of Accuracy and Precision in quantitative IVD For in-vitro diagnostic use Use by/Expiration Date 327 001 (1 vial ) 1 x 5 ml
LOT Batch Code/Lot number CAUTION. Consult instructions 327 002 (5 vials) 5 x 5 ml
In vitro Diagnostics REF Catalogue Number for use
REF: 327 001 (1 vial ) 1 x 5 ml

REF: 327 002 (5 vials) 5 x 5 ml
Description
Expected Values
SP-PATHOTROL is a lyophilized human-based control serum.
1. The expected value of specific assays are provided on the
Storage and stability assay value sheet accompanying each kit, and are lot specific.
Unopened bottles must be stored at 2 - 8°C and are stable up to
the expiration date printed on the labels 2. The expected values are obtained using replicate assay of
each manufactured lot of SP-Pathotrol.
After reconstitution the serum can be used , stored at 2 - 8°C and
protected from light , in between 5 days. 3. The individual laboratory values should fall within the expected
Bilirubin is stable 1 day ,refrigerated . values.
The frozen serum is stable for at least 30 days at < -20°C .
4. It must however be noted that each laboratory should establish
Warnings and Precautions its own normal values and reference range according to GLP.
Each individual blood donation used for this serum was found
negative when tested with FDA-approved methods on HBsAg, anti-
Literature
HIV 1+2 and anti-HCV. Nevertheless the serum should be treated
for safety reasons always as a potentially infectious material . 1. Röhle G, Siekmann L. Quality assurance of quantitative
determination. In: Thomas L, editor. Clinical laboratory
Preparation diagnostics.1st ed. Frankfurt:TH-Books Verlagsgesellschaft;
Reconstitute carefully by adding exactly 5 ml of distilled water. 1998. p. 1393-1401
Allow the vial to stand for 30 minutes ,swirling occasionally.
Avoid foaming i.e. do not shake! 2. Biosafety in Microbiological and Biomedical Laboratories. U.S.
For alkaline phosphatase the control serum should stand for 2 Department of Health and Human Services, Washington 1993
hours at +25 °C before use . (HHS Publication No. [CDC] 93-8395).
Procedure 3. Richtlinien der Bundesärztekammer zur Qualitätssicherung in

Refer to the package inserts of the reagent kits medizinischen Laboratorien. Deutsches Ärzteblatt 1988;85:
B519–B532.
Assay Values and Ranges
Assay values for analytes for which approved reference methods 4. Richtlinie der Bundesärztekammer zur Qualitätssicherung
are available were determined according to Guidelines of the quantitativer laboratoriumsmedizinischer Untersuchungen
German Federal Medical Council [Bundesaerztekammer] from 1987 Deutsches Ärzteblatt 2002;98:A 2747-59.
(reference method values) [3].
Ranges of acceptance were calculated as assigned value ± three
times the maximum tolerable deviation of a single value according to
the Guidelines of the German Federal Medical Council [4].
158 159
SP-MULTICAL SYMBOLS IN PRODUCT LABELLING ORDERING INFORMATION
Multiparameter calibration serum for the EC REP Authorised Representative Temperature Limitation CATALOG NO QUANTITY
standradization of quatitative tests on IVD For in-vitro diagnostic use Use by/Expiration Date 328 001 (1 vial ) 1 x 5 ml
328 001 (5 vials) 5 x 5 ml
clinical chemical Analytes REF Catalogue Number for use
REF: 328 001 (1 vial ) 1 x 5 ml

REF: 328 002 (5 vials) 5 x 5 ml
Description Preparation
SP-MULTICAL is an analyzed calibration material based on The lyophilisate is vacuum sealed; therefore the vial should be
lyophilized human serum. reconstituted carefully with exactly 3 ml of distilled water.
Close the vial carefully and allow the calibrator to stand for 30
Storage and stability minutes swirling occasionally.
Unopened bottles have to be stored in refrigerator at 2 – 8 °C and Avoid foaming! Do not shake!
are stable up to the expiry date printed on the labels.
Procedure
After reconstitution the tightly closed calibrator can be used within Refer to the package inserts of the reagent kits
the times and temperatures given below:
Expected Values
General:
18 – 25 °C => 8 hours 1. The expected value of specific assays are provided on the
2 – 8 °C => 2 days assay value sheet accompanying each kit, and are lot specific.
< – 25 °C => 30 days
2. The expected values are obtained using replicate assay of
Bilirubin (in the dark) each manufactured lot of SP-Multical.
18 – 25 °C => 4 hours
2 – 8 °C => 8 hours 3. The individual laboratory values should fall within the expected
< – 25 °C => 14 days values.
Acid Phosphatase 4. It must however be noted that each laboratory should establish
(Total and Prostatic) its own normal values and reference range according to GLP.
18 – 25 °C => 4 hours
2 – 8 °C => 8 hours
Literature
< – 25 °C => 14 days
1. Dati F. Reference materials and guidelines for standardization
- freeze only once of methods in laboratory medicine. In: Thomas L, editor.
Clinical laboratory diagnostics. 1st ed. Frankfurt: TH-Books
Warnings and Precautions Verlagsgesellschaft; 1998. p. 1404-26.
Each individual blood donation used for production of
SP- MULTICAL. was found to be non-reactive when tested with 2. Moss DW, Henderson AR. Enzymes . In: Burtis CA,Ashwood
approved methods for HBsAg, anti-HIV 1+2 and anti-HCV. ER, editors. Tietz Textbook of Clinical Chemistry. 2nd ed.
Philadelphia: W.B SaundersCompany; 1994. p. 735-896.
As there is no possibility to exclude definitively that products derived
from human blood transmit infectious agents, it is recommended to 3. Biosafety in Microbiological and Biomedical Laboratories. U.S.
handle the calibrator with the same precautions as valid for patient Department of Health and Human Services, Washington 1993
specimens. (HHS Publication No. [CDC] 93-8395)
160 161
HbA1c Calibrator
For the Quantitative Determination of HbA1c
REF: 356 001 1 X 0.5 ml
Intended use
Calibration Blood for standardization of quantitative determinations 3. Reagents containing sodium azide must be handled with
caution: Do not swallow or allow to contact skin or mucous
of HbA1c
membranes! Sodium azide can form explosive azides when
contacting metals such as copper or lead.
Composition
Assigned Values
HbA1c Calibrator is a lyophilisate from hemolyzed human blood,
stabilized. The given value is valid only for the Spectrum colorimetric HbA1c
Preservative = 0.095 % sodium azide. The value of HbA1c is in the test, and only for the given specific lot no.
normal range. The value has been assigned to be traceable to corresponding
reference materials.
Assigned Value: Stated on HbA1c Calibrator Label.
The expiry date of the product in the unopened vials, stored at
2 - 8 oC, is printed on the labels. Procedure
Preparation Refer to the package inserts of the reagent kits respectively

instrument procedures.
Reconstitute with 0.50 ml of dist. water - swirl gently and let stand
for about 15 min - the blood is ready for use. Waste Management
It has to be stored at 2 - 8°C and is stable for 2 weeks . It can be Please refer to the local laws and their requirements
used for at least 3 more months at < - 20°C if stored tightly closed
Literature
1. Dati F. Reference materials and guidelines for standardization
1. For In Vitro Diagnostic use only. of methods in laboratory medicine. In: Thomas L, editor.
Clinical laboratory diagnostics. 1st ed. Frankfurt: TH-Books
2. Each individual donation intended for use in manufacture of Verlagsgesellschaft; 1998. p. 1404-26.
the blood was tested for hepatitis B surface antigen (HBsAg),
anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV 2 by 2. Biosafety in Microbiological and BiomedicalLaboratories. U.S.
FDA approved tests. Only donations with negative findings Department of Health and Human Services, Washington 1993
were used. Nevertheless each product of human source
should be handled with appropriate care in accordance with ORDERING INFORMATION
recommended procedures for biohazardous materials, since
absence of infectious agents can never be proven. CATALOG NO QUANTITY
256 001 1 X 0.5 ml
162 163
C-immunochemistry
164 165
Antistreptolysin O Titre (ASOT) SYMBOLS IN PRODUCT LABELLING Results and Interpretation References
A rapid latex slide test for the detection of EC REP Authorised Representative Temperature Limitation
antistreptolysin O antibodies in serum IVD For in-vitro diagnostic use Use by/Expiration Date Negative result: 1. Alouf et al. Biochimie 1973 ; 56-61.
No agglutination of the latex particles suspension within two minutes.
2. Ginsburg, I. (1972). J. Infect.. Dis., 126, 294-340.
Positive result:
REF: 510 001 50 test (Complete Kit) An agglutination of the latex particles suspension will occur within 3. Halbert, SP. Ann. N.Y. Acad. Sci., 103, 1027:1051; 1963.
REF: 510 002 100 test (Copmlete Kit) two minutes, indicating an ASO level of more than 200 IU/ml.
4. Klein GL, Applied Microbiology, 21:999, 1971.
REF: 510 003 50 test (latex with positive control) Semi-Quantitative Test
REF: 510 004 100 test (latex with positive control) Slides and Stirrers. 5. Klein GC: Manual of Clinical Immunology ASM 264-273:1976.
1. Serum to be titrated is serially diluted (1:2, 1:4, 1:8 etc) in 0.9
REF: 510 000 100 test (latex only) NOTE: Negative Control Serum, Slides and Stirrers are only g/L saline solution. 6. Rantz LD, DiCapri JM, Randall E. Am. J. Med. Sci., 24,1952.
REF: 510 005 200 test (latex only) included in Complete Kits REF: 510 001 (50 test) &
REF: 510 002 (100 test) 2. Place one drop of positive control on slide. Do not attempt to 7. Schmidt et al. Rheumatol. 1970 ; 29 : 29-32.
dilute the ASO positive control serum for comparative or other
Intended Use Storage & Stability purposes as no correlation exists between actual titre of the
Rapid latex agglutination test for the qualitative screening and The reagents are stable up to the expiration date specified when control and titre of unknown sera.
semi-quantitative determination of antistreptolysin O ( ASO) stored at 2 – 8 C.
antibodies in human serum. 3. Place 50 ml of each serum dilution individually in successive CATALOG NO QUANTITY
Precautions and Warnings circles on the slide and proceed as in screening methodology. 510 001 50 test
Background All human blood components used to prepare controls have been 510 002 100 test
In infections caused by ß-hemolytic streptococci, Streptolysin O is tested for Hepatitis B surface antigen (HBsAg) and HTLV-III an- Results and Interpretation
liberated from the bacteria stimulating production of antistreptolysin tibodies by FDA approved procedure and found to be non-reactive. The serum ASO titre can be defined as the highest dilution showing 510 003 50 test
O (ASO) antibodies. The extent and degree of the infection can be No known test method for HBsAg or HTLV-III antibodies offers total a positive result. The approximate ASO level (IU/ml) present in the 510 004 100 test
monitored by measuring the levels of these antibodies. Increase in assurance that a human derived product will not transmit hepatitis or sample can be optained by the following formula:
ASO titre generally occurs one to four weeks after onset of infection HTLV-III virus. The user is therefore cautioned to handle reagents as 510 000 100 test
with b-hemolytic streptococci Group A. As the infection subsides, the if being capable of transmitting these diseases. ASO Titre (IU/ml) = Highest dilution with positive reaction x 510 005 200 test
titre declines and returns to normal levels within six months. If the Reagent sensitivity ( 200 IU/ml)
titre does not decrease, a recurrent or chronic infection may exist. Specimen Collection and Preservation
Use only serum specimens , plasma samples are not suitable for the e.g. if the agglutination is present up to a titre 1:8 , the approximate
Test Principle o
test. Serum samples can be stored for 24 hrs at 2 – 8 C, for longer serum ASO level is 8 x 200 = 1600 IU/ml
o
Spectrum ASO latex reagent is a suspension of polystyren particles Storage it is recommended to store the samples at -20 C
sensitized with streptolysin O. When the latex reagent is mixed with Expected Value
a serum containing antibodies to streptolysin O, visible agglutination Procedure Up to 200 IU/ml
occurs. The latex reagent has been produced so that agglutination
will take place only when the level of antibodies to streptolysin O is Qualitative Test (Screening) Limitations of the Procedure
greater than 200 IU/ml. Occasional agglutinations observed after 4 minutes have no
1. Bring all reagents and specimens to room temperature. diagnostic significance.
Reagents Highly haemolyzed and lipemic serum as well as plasma interfere
Spectrum ASO latex kit contains the following reagents: 2. Place one drop (˜ 50 ml) of the positive control and 50 ml of the with the test.
patient serum into separate circles on the glass slide.
Latex Reagent (bottle with green cap):
A suspension of polystyrene latex particles in glycine-saline buffer 3. Shake the ASO latex reagent gently and add one drop (45 ml)
pH: 8.6 ± 0.1, coated with streptolysin O. on each circle next to the sample to be tested and control.
Positive Control Serum (bottle with red cap): 4. Mix well using disposable stirrer spreading the mixture over
Is prepared from a stabilized human serum pool containing more the whole test area and tilt the slide gently. Agitate for about 2
than 200 IU/ml antistreptolysin O. Both reagents contain 0.9 g/L minutes with rotator or by hand and observe for the
Sodium azide as a preservative. presence or abscence of agglutination.
Negative Control Serum (bottle with white cap):

Reagent contain 0.9 g/L Na azide as a preservative.
166 167
C Reactive Protein (CRP) SYMBOLS IN PRODUCT LABELLING Results and Interpretation References
A rapid latex slide test for the EC REP Authorised Representative Temperature Limitation Negative result: No agglutination of the latex particles suspension
For in-vitro diagnostic use Use by/Expiration Date
detection of CRP in serum IVD
within two minutes. 1. Bowman BH. In:Hepatic Plasma Protein. San Diego: Academic
Positive result: An agglutination of the latex particles suspension Press;1993:47-95.
will occur within two minutes, indicating a CRP level of more than
6 mg/L. 2. Halbert, SP. Ann. N.Y. Acad. Sci., 103, 1027:1051; 1963.
REF: 514 001 50 test (Complete Kit)
REF: 514 002 100 test (Complete Kit) Semi-Quantitative Test 3. Klein GL, Applied Microbiology, 21:999, 1971.
REF: 514 003 50 test (latex with positive control) 1. Serum to be titrated is serially diluted (1:2, 1:4, 1:8 etc) in 0.9 4. Klein GC: Manual of Clinical Immunology ASM 264-273: 1976.
REF: 514 004 100 test (latex with positive control) Slides g/L saline solution.
5. Pepys MB et al. Lancet 1961 ; 1 : 653-660.
REF: 514 000 100 test (latex only) NOTE: Negative Control Serum, Slides are only 2. Place one drop of positive control on slide. Do not attempt to
REF: 514 005 200 test (latex only) included in Complete Kits REF: 514 001 (50 test) & dilute the CRP positive control serum for comparative or other 6. Rantz LD, DiCapri JM, Randall E. Am. J. Med. Sci., 24, 1952.
REF: 514 002 (100 test) purposes as no correlation exists between actual titre of the
control and titre of unknown sera.
Intended Use Storage & Stability
Rapid latex agglutination test for the qualitative screening and semi- The reagents are stable up to the expiration date specified when 3. Place 50 ml of each serum dilution individually in successive
quantitative determination of C Reactive Protein (CRP) in human
o
stored at 2 – 8 C. circles on the slide and proceed as in screening methodology. CATALOG NO QUANTITY
serum. 514 001 50 test
Precautions and Warnings Results and Interpretation 514 002 100 test
Background All human blood components used to prepare controls have been The serum CRP titre can be defined as the highest dilution showing
Tissue-damaging associated with inflammatory diseases, infection tested for Hepatitis B surface antigen (HBsAg) and HTLV-III an- a positive result. The approximate CRP level ( mg/L) present in the 514 003 50 test
and neoplasms are associated with a major accute phase response tibodies by FDA approved procedure and found to be non-reactive. sample can be optained by the following formula: 514 004 100 test
of the C-reactive protein ( CRP ) and other acute phase reactants. No known test method for HBsAg or HTLV-III antibodies offers total
The CRP response frequently precedes clinical symptoms, including assurance that a human derived product will not transmit hepatitis or CRP Titre ( mg/L) = Highest dilution with positive reaction x 514 000 100 test
fever. Measuring changes in the concentration of CRP provides HTLV-III virus. The user is therefore cautioned to handle reagents as Reagent sensitivity ( 6 mg/lL) 514 005 200 test
useful diagnostic information about how acute and how serious a if being capable of transmitting these diseases.
disease is. It also allows the assessment of complications during the e.g. if the agglutination is present up to a titre 1:8, the approximate
disease and judgement about the disease genesis. Specimen Collection and Preservation serum CRP level is 8 x 6 = 48 mg/L.
Use only serum specimens, plasma samples are not suitable for the
Test Principle o
test. Serum samples can be stored for 24 hrs at 2 – 8 C, for longer Expected Value
o
Spectrum CRP latex reagent is a suspension of polystyrene parti- storage it is recommended to store the samples at -20 C. Up to 6 – 8 mg/L.
cles sensitized with anti-human CRP. When the latex reagent is
mixed with a serum containing C-reactive protein, visible agglut- Procedure Limitations of the Procedure
ination occurs. The latex reagent has been produced so that agglu- Occasional agglutinations observed after 4 minutes have no
tination will take place only when the level of CRP is greater than 6 Qualitative Test (Screening) diagnostic significance.
mg/L.
1. Bring all reagents and specimens to room temperature. Highly haemolyzed and lipemic serum as well as plasma interfere
Reagents with the test.
2. Place one drop (50 ml) of the positive control and 50 ml of the
Spectrum CRP latex kit contains the following reagents: patient serum into separate circles on the glass slide.
Latex Reagent (bottle with green cap): 3. Shake the CRP latex reagent gently and add one drop(45 ml)
A suspension of polystyrene latex particles in glycine-saline buffer on each circle next to the sample to be tested and control.
pH: 8.6 ± 0.1, coated with anti-human CRP antibodies.
4. Mix well using disposable stirrer spreading the mixture over
Positive Control Serum (bottle with red cap): the whole test area and tilt the slide gently. Agitate for about 2
Is prepared from a stabilized human serum pool containing CRP minutes with rotator or by hand and observe for the presence
as an antigen. or abscence of agglutination.
Both reagents contain 0.9 g/L Sodium azide as a preservative.
Negative Control Serum (bottle with white cap):

168 169
Rheumatoid Factor (RF) SYMBOLS IN PRODUCT LABELLING
Semi-Quantitative Test ORDERING INFORMATION
A rapid latex slide test for the detection EC REP Authorised Representative Temperature Limitation 1. Serum to be titrated is serially diluted (1:2, 1:4, 1:8 etc) in 0.9 CATALOG NO QUANTITY
of Rheumatoid Factor in serum IVD For in-vitro diagnostic use Use by/Expiration Date g/L saline solution. 518 001 50 test
2. Place one drop of positive control on slide. Do not attempt to 518 002 100 test
dilute the RF positive control serum for comparative or other
REF: 518 001 50 test (Complete Kit) purposes as no correlation exists between actual titre of the 518 003 50 test
REF: 518 002 100 test (Complete Kit) control and titre of unknown sera. 518 004 100 test
3. Place 50 ml of each serum dilution individually in successive
REF: 518 003 50 test (latex with positive control) circles on the slide and proceed as in screening methodology. 518 000 100 test
REF: 518 004 100 test (latex with positive control) Storage & Stability
The reagents are stable up to the expiration date specified when Results and Interpretation
o
REF: 518 000 100 test (latex only) stored at 2 – 8 C. The serum RF titre can be defined as the highest dilution showing
a positive result. The approximate RF level ( IU/ml) present in the
Precautions and Warnings sample can be optained by the following formula:
Intended Use All human blood components used to prepare controls have been CRP Titre (mg/L) = Highest dilution with positive reaction x
Rapid latex agglutination test for the qualitative screening and tested for Hepatitis B surface antigen (HBsAg) and HTLV-III an-ti- Reagent sensitivity (10 IU/ml)
semi-quantitative determination of rheumatoid factor (RF) in human bodies by FDA approved procedure and found to be non-reactive.
serum. No known test method for HBsAg or HTLV-III antibodies offers total e.g. if the agglutination is present up to a titre 1:8, the approximate
assurance that a human derived product will not transmit hepatitis or serum RF level is 8 x 10 = 80 IU/ml.
Background HTLV-III virus. The user is therefore cautioned to handle reagents as
Rheumatoid factors are immunoglobulins which are directed against if being capable of transmitting these diseases. Expected Value
the Fc portion of IgG. Rheumatoid arthritis is a chronic systemic Not clearly specified . However , it has been found that the existence
disease of unknown etiology. Its diagnosis is based on combined Specimen Collection and Preservation of significantly high titre ( more than 30 IU/ml ) are present in more
clinical and radiographic analysis. The determination of RF is the Use only serum specimens, plasma samples are not suitable for the than 70 % of patients with rheumatoid arthritis.
o
laboratory test that is most commonly used not only for the diagnosis test . Serum samples can be stored for 24 hrs at 2 – 8 C, for longer
of rheumatoid arthritis but also assists in the prognosis of the disease storage it is recommended to store the samples at -20 C.
o
Limitations of the Procedure
and in the monitoring of therapeutic response. Occasional agglutinations observed after 4 minutes have no
Procedure diagnostic significance. Highly haemolyzed and lipemic serum as
Test Principle well as plasma interfere with the test.
Qualitative Test (Screening)
Spectrum RF latex reagent is a suspension of polystyrene particles
sensitized with human gamma globulin. When the latex reagent is 1. Bring all reagents and specimens to room temperature.
mixed with a serum containing rheumatoid factor, visible agglutination 2. Place one drop (50 ml) of the positive control and 50 ml of the
References
occurs. The latex reagent has been produced so that agglutination patient serum into separate circles on the glass slide. 1. Ball J. et al. Ann Rheum. Die 1963 ; 22 : 311-314.
will take place only when the level of RF is greater than 10 IU/ml . 3. Shake the RF latex reagent gently and add one drop (45 ml) on 2. Halbert, SP. Ann. N.Y. Acad. Sci., 103, 1027:1051; 1963.
each circle next to the sample to be tested and control.
3. Klein GL, Applied Microbiology, 21:999, 1971.
Reagents 4. Mix well using disposable stirrer spreading the mixture over
Spectrum RF latex kit contains the following reagents: the whole test area and tilt the slide gently. Agitate for about 2 4. Klein GC: Manual of Clinical Immunology ASM 264-273:1976.
minutes with rotator or by hand and observe for the presence 5. Rantz LD, DiCapri JM, Randall E. Am. J. Med. Sci., 24,1952.
or abscence of agglutination.
Latex Reagent (bottle with green cap):
A suspension of polystyrene latex particles in glycine-saline buffer
pH: 8.6 ± 0.1, coated with human gama globulin. Results and Interpretation
Negative result: No agglutination of the latex particles suspension
Positive Control Serum (bottle with red cap): within two minutes.
Is prepared from a stabilized human serum pool containing RF.
Both reagents contain 0.9 g/L Sodium azide as a preservative. Positive result: An agglutination of the latex particles suspension
will occur within two minutes, indicating a RF level of more than
Negative Control Serum (bottle with white cap): 10 IU/ml.
Slides and Stirrers
NOTE: Negative Control Serum, Slides and Stirrers are only

included in Complete Kits REF: 518 001 (50 test) &
REF: 518 002 (100 test)
170 171
D- turbidimetric immunoassay
172 173
Quality Control
Antistreptolysin O (ASO) SYMBOLS IN PRODUCT LABELLING Control sera are recommended to monitor the perfomance of manual
EC REP Authorised Representative Temperature Limitation and automated assay procedures. Each laboratory should establish
Immuno-Turbidimetry IVD For in-vitro diagnostic use Use by/Expiration Date its own Quality Control Scheme and corrective actions if controls do
LOT Batch Code/Lot number CAUTION. Consult instructions not meet the acceptable tolerances.
REF: 596 001 100 test Expected Values
R1 Buffer 2 X 20 ml Normal values 0 - 200 IU/ml
R2 Latex 1 X 6 ml
Each laboratory should establish an expected range for the
geographical area in which it is located.
References
Intended Use ASO Standard: 1. Tadzynsky LA, Ryan ME. Diagnostic of rheumatoid fever. A
The Standard is stable to the expiration date on the vial label when guide to criterial and manifestations. Postgrad Med 1986;
In vitro diagnostic reagents for the quantitative determination of capped and stored at (2 - 8 ºC). 79:295.
Antistreptolysin O (ASO) in human serum by means of particle- 2. Dillon,H.C.jr,Reeves M.A,aM.J.Med,56,333-346(1974).
enhanced turbidimetric immunoassay. Once opened the standard is stable for 6 weeks if stored tightly 3. Bach GL, Cadotte R, Wiatr RA, et al. Latex antiestreptolysin O
closed at 2 - 8 ºC after use. test as a tube dilution procedure. Am J Clin Pathol 1972; 57:
Background 209.
For further storage at - 30 ºC divide standard into aliquots. Stability 4. Klein,G.C.Baker,C.N,Jones,W.L,21,999-1001(1971).
Immunological testing for specific antibodies to streptococcal 3 months . once thawed never freeze again.
metabolites provides important information regarding a prior 5. Curtis GDW, Kraak WAG, Mitchell RG. Comparison of latex
streptococcal infection. Antibodies are formed against both the and hemolysis tests for determination of antiestreptolysin O
Specimen Collection and Preparation (ASO) antibodies. J Clin Pathol 1988; 41: 1331.
pathogen itself and its metabolic products. An example for the
Serum specimens should be collected by venipuncture following
latter is the antibody against streptolysin O, an enzyme secreted by
good laboratory practices. Suitable assay specimens are human
betahaemolytic streptococci of the Landfield Group A.
serum samples, as fresh as possible (stored up to 2 days at 2 - 8 ºC)
Antistreptolysin O (ASO) testing is thus used for the diagnosis of
or deep-frozen. Any additional clotting or precipitation which occurs
non suppurative complications of the infections caused by these
due to the freeze/thaw cycle should be removed by centrifugation
pathogens: acute rheumatic fever or acute poststreptococcal
prior to assay.
glomerulonephritis. In the determination of antibodies to various ORDERING INFORMATION
Heavily lipemic sera may lead to a non-specific reaction due to
streptococcal exoenzymes preference is to be given to anti-
chylomicrons. Lipemic specimens, or turbid frozen specimens CATALOG NO QUANTITY
streptolysin O, since this sensitive parameter is found to be elevated
after thawing, must be clarified before the assay by high-speed
in about 80 to 85% of cases. 596 001 100 test
centrifugation (15 min at approx. 15.000 rpm).
Test Principle Procedure

This ASO test is based upon the ASO antigen-antibody reaction. 1. Bring the reagents and the photometer to 37ºC
2. Assay conditions:
Temperature 37ºC
R1 Buffer Cuvette 1cm light path
Phosphate buffered saline (pH 7.43)
3. Adjust the instrument to zero with distilled water .
Enhancer.
4. Pipette into a cuvette :
Sodium azide (0.095 g/L)
Standard Sample
R2 Latex reagent Reagent (R1) 400 ml 400 ml
Glycine Buffer (pH8.2)
Standard 5ml ------
ASL senesitized Latex (0.17 %)
Sodium azide 0.95 g/L. Sample ------ 5ml
Materials required but not provided with the kit 5. Mix and incubate for 2 minutes,read absorbance (A1)
Reagent (R2) 60ml 60ml
1- Standard
ASO concentration is stated on the vial label. After addition of R2, incubate and after 5 minutes record 2nd
reading (A2)
2-Controls
Precautions and Warnings Calculation

For in vitro diagnostic use only. Do not pipette by mouth. Reagents Generate a reference curve by successive 1 : 2 dilutions of standard
containing sodium azide must be handled with precaution. Sodium in saline (At Least 4 points are recommended). Use Saline as zero
azide can form explosive azides with lead and copper plumbing.
point. Determine D absorbance of the sample and each standard as
Since absence of infectious agents cannot be proven, all specimens
and reagents obtained from human blood should always be handled following:
with precaution using established good laboratory practices. D absorbance of sample = (A2 - A1) sample
Disposal of all waste material should be in accordance with local D absorbance of each standard = (A2 - A1) for each standard
guidelines.
Plot the calibration curve and obtain the result.
As with other diagnostic tests, results should be interpreted
considering all other test results and the clinical situation of the
patient. Sensitivity
10.0 IU/mL.
Reagent Preparation,Storage and Stability
All reagents are supplied ready to use. Linearity
Reagents in the original vial are stable to the expiration date on the 400 IU/mL.
vial label when capped and stored at (2 - 8 ºC).
174 175
Antistreptolysin O (ASO) SYMBOLS IN PRODUCT LABELLING
Reagent Preparation and Stability
Working Reagent is prepared with 1 part of Latex Reagent and 4
interferences
Hemoglobin (10 g/L) , bilirrubin (20 mg/dL) and lipemia (10 g/L) ,and
Turbi Latex EC REP Authorised Representative
parts of Diluent. Prepare a fresh WR based on its workload. Shake
gently the reagents before pipetting.
rheumatoid factors (600 IU/ml) do not interfere.Other substances
may interfere6.
e.g. 400 ml Diluent + 100 ml Latex Reagent.
REF Catalogue Number for use References
REF: 559 001 100 test 1. Tadzynsky LA, Ryan ME. Diagnostic of rheumatoid fever. A
R1 Buffer 2 X 20 ml stability : 1 month at at 2 - 8 ºC. guide to criterial and manifestations. Postgrad Med 1986;
R2 Latex 1 X 10 ml 79:295.
ASO Calibrator: Reconstitute with 1 ml distelled water. mix gently 2. Bach GL, Cadotte R, Wiatr RA, et al. Latex antiestreptolysin O
and incubate at room temperature for 10 minutes before use. test as a tube dilution procedure. Am J Clin Pathol 1972; 57:
209.
Intended Use Precautions and Warnings Stability: 1 month at at 2 - 8 ºC or 3 months at at -20 ºC 3. Rantz LA, Randall E. A modification of the technic for
In vitro diagnostic reagents for the quantitative determination of For in vitro diagnostic use only. Do not pipette by mouth. Reagents determination of the antiestreptolysin titer. Proc Soc Exp Biol
Antistreptolysin O (ASO) in human serum by means of particle- containing sodium azide must be handled with precaution. Sodium Quality Control Med 1945; 59:22.
Control sera are recommended to monitor the perfomance of manu-
enhanced turbidimetric immunoassay. azide can form explosive azides with lead and copper plumbing. 4. Curtis GDW, Kraak WAG, Mitchell RG. Comparison of latex
al and automated assay procedures. Each laboratory should estab-
Since absence of infectious agents cannot be proven, all specimens lish its own Quality Control Scheme and corrective actions if controls and hemolysis tests for determination of antiestreptolysin O
Background and reagents obtained from human blood should always be handled do not meet the acceptable tolerances. (ASO) antibodies. J Clin Pathol 1988; 41: 1331.
Immunological testing for specific antibodies to streptococcal with precaution using established good laboratory practices. 5. Passing H, Bablok W. A new biometrical procedure for testing
metabolites provides important information regarding a prior Procedure the equality of measurements from two analytical methods.
streptococcal infection. Antibodies are formed against both the Disposal of all waste material should be in accordance with local 1. Bring the reagents and the photometer to 37ºC Application of linear regression procedures for method
pathogen itself and its metabolic products. An example for the guidelines. 2. Assay conditions: comparison studies. Part I. J Clin Chem Clin Biochem 1983;
latter is the antibody against streptolysin O, an enzyme secreted by Wavelength 540 nm (530 -550 nm) 21:709-20.
betahaemolytic streptococci of the Landfield Group A. As with other diagnostic tests, results should be interpreted Temperature 37ºC 6. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
Antistreptolysin O (ASO) testing is thus used for the diagnosis of considering all other test results and the clinical situation of the Cuvette 1cm light path 207 . 224.
non suppurative complications of the infections caused by these patient. 3. Adjust the instrument to zero with distilled water .
pathogens: acute rheumatic fever or acute poststreptococcal 4. Pipette into a cuvette : ORDERING INFORMATION
glomerulonephritis. In the determination of antibodies to various Storage and Stability Working Reagent 500 ml CATALOG NO QUANTITY
streptococcal exoenzymes preference is to be given to anti- Reagents in the original vial is stable to the expiration date on the vial Calibrator or Sample 5 ml 559 001 100 test
streptolysin O, since this sensitive parameter is found to be elevated label when capped and stored at (2 - 8 ºC). Immediately following the
in about 80 to 85% of cases. completion of an assay run, the reagent vial should be capped until 5.Mix and read absorbance immediately (A1) and after 2
next use in order to maximize curve stability. Do not freeze reagents. min (A2) of the sample addition.
Test Principle The ASO latex reagent should have a white, turbid appearance free
The present ASO test is based upon the reactions between of granular particulate. Visible agglutination or precipitation may be Calculation
antibodies against streptolysin O (ASO) and latex particles bound a sign of deterioration, and the reagent should be discarted.
(A2-A1) sample
streptolysin O. ASO values are determined photometrically. x Calibrator concentration= IU/ml ASO
The ASO diluent should be clear and colourless. Any turbidity may (A2-A1) calibrator
Reagents be sign of deterioration and reagent should be discarted.
R1 Diluent Sensitivity
Trisbuffer 20mmol/L,pH8.2.Sodium azide0.95 g/L. Specimen Collection and Preparation Up to 20 IU/mL.
R2 Latex reagent good laboratory practices. Suitable assay specimens are human Linearity
Latex particles coated with streptolysin O,pH 10.0 serum samples, as fresh as possible (stored up to 2 days at 2 - 8 ºC) Up to 800 IU/mL.
Sodium azide 0.95 g/L. or deep-frozen. Any additional clotting or precipitation which occurs
due to the freeze/thaw cycle should be removed by centrifugation specimens showing higher concentration should be diluted 1+2
Calibrator prior to assay. using physiological saline and repeat the assay (result×3).
Human serum.ASO concentration is stated on the vial label. Heavily lipemic sera may lead to a non-specific reaction due to
chylomicrons. Lipemic specimens, or turbid frozen specimens Expected Values
All raw materials of human origin used in the manufacture of this after thawing, must be clarified before the assay by high-speed Normal values < 200 IU/ml (adults) and 100 IU/ml (children<5 years
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 centrifugation (15 min at approx. 15.000 rpm). old).
and HCV with commercially available test methods. However,
this product should be handled as though capable of transmitting Each laboratory should establish an expected range for the
infectious diseases geographical area in which it is located.
176 177
ASO Standard Super High SYMBOLS IN PRODUCT LABELLING ASO Control SYMBOLS IN PRODUCT LABELLING
EC REP Authorised Representative Temperature Limitation EC REP Authorised Representative Temperature Limitation
REF 345 001 1x1 ml IVD For in-vitro diagnostic use Use by/Expiration Date REF 340 001 1 x 1 ml IVD For in-vitro diagnostic use Use by/Expiration Date
LOT Batch Code/Lot number CAUTION. Consult instructions LOT Batch Code/Lot number CAUTION. Consult instructions
REF Catalogue Number for use REF Catalogue Number for use
Consult instructions for use Manufactured by Consult instructions for use Manufactured by
Specifications Intended Use

Accuracy control for the determination of ASO in serum by
turbidimetry and nephelometry
Lot number : ASO 3014
Composition
Expiration Date : June 2016 A dilution of humen plasma containing high levels of ASO with
phosphsate buffered saline.
Source : Human plasma Preservative: 0.095 g% sodium azide.
o
Storage: 2 - 8 C Do Not Freeze Storage and Stability
The expiry date of the product at +2 to +8 ºC is listed on the label.
Stability after opening the vial : 6 weeks After first opening the control can be used for 6 weeks if stored
tightly closed at +2 to+8 ºC after use.
Do not freeze.

1. For “In Vitro Diagnostic Use”
2. Each individual donation intented for use in manufacturing
of protein control serum was tested for hepatities B surface
antigin (HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1
and HIV2 by FDA required test. Only donations with negative
findings were used for its manufacture.Nevertheless every
product obtained from human body fluids should be handled
with appropriate care in accordance with recommended
procedures for biohazardous materials since absence of
infectious agents can never be proven .
3. Reagents containing sodium azide must be handled with
due caution: Do not ingest or allow to contact skin or mucous
membranes !
Assigned Value
LOT No. : ASOT 3014
ASO : 188 ( 173 - 203 ) IU/mL.
Target ASO IU/mL

ASO Standard Super High
419
CATALOG NO QUANTITY
340 001 1 X 1 ml
178 179
b2 -MICROGLOBULIN (b2 -m) SYMBOLS IN PRODUCT LABELLING Incubation time 4 min. establish its own reference intervals.
EC REP Authorised Representative Temperature Limitation Procedure
IVD For in-vitro diagnostic use Use by/Expiration Date The reagents are ready to use as supplied. Latex reagent should be References
REF: 553 001 100 test gently shaken (invert the recipient 3-4 times) before each use. 1. F. Ortega y col.. Influencia pronóstica de la b2-microglobulina
R1 Buffer : 25 ml en el mieloma múltiple. Med. Clin. (Barc) 1992; 99: 645-648
R2 Latex : 5 ml Volume R1/Buffer reagent: 250 ml
2. Lifson A.R, Hessol N A, Buchbinder S P, O.Malley P M,
Volume R2/Latex reagent: 50 ml Barnhart L, Segal M, Katz M H, Holmberg S D. Serum ß2-
Volume sample: 2 ml microglobulin and prediction of progression to AIDS in HIV
Intended Use Precautions and Warnings infection. The Lancet 1992; 339: 1436-40.
In vitro diagnostic reagents for the quantitative determination of For in vitro diagnostic use only. Do not pipette by mouth. Reagents Step 1: mix R1 and R2, add sample and read 1st reading immediately 3. Weissel M, Scherak O, et al. Serum b2-microglobulin and
b2 -Microglobulin (b2 -m) in serum and urine by means of particle containing sodium azide must be handled with precaution. Sodium after mixing. SLE. Arthritis Rheum 1976; 19:986.
enhanced turbidimetric immunoassay. azide can form explosive azides with lead and copper plumbing. 4. Evrin P E, Paterson P A, Wide L, Berggard Y.
Since absence of infectious agents cannot be proven, all specimens Step 2: after 4 min read 2nd reading. Radioimmunoassay of ƒÒ2- microglobulin in human biological
Background and reagents obtained from human blood should always be handled fluids. Scand J clin Lab Invest 1971; 28: 439-43.
b2-microglobulin (b2-m) , an 11,800-Da protein consisting of 100 with precaution using established good laboratory practices. Note: Volume, time and wavelength are recommended. Adjust them 5. Schardijn G, Statius van Eps L. b2-microglobulin: Its
aminoacid residues, first isolated from urine by Berggard and depending of analyser features. significance in the evaluation of renal function. Kidney
International 1987; 32: 635 41.
Bearn, is located on the surface of lymphocytes and other nucleated Disposal of all waste material should be in accordance with local
cells and was identified to be the light chain of the class I major guidelines. This reagent is intended to be used in clinical chemistry analysers. 6. Becker W. Die Standardddidierung immunchemischer
Plasmaprotein- Bestimmungen. Laboraatoriummblätter 1980;
histocompatibility complex. The cell membrane turnover is the main Adaptations for some of them are available.
30: 25 . 32.
source of b2-m in serum, where its level rises when its production As with other diagnostic tests, results should be interpreted
rate increases. Free b2-m is filtered by the glomerulus and considering all other test results and the clinical situation of the Calibration and Quality Control 7. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
207 .224.
subsequently reabsorbed almost completely in the proximal tubular patient. Standardization: use Spectrum Calibrators. The method was
cells, where it is catabolized. Increased urinary excretion of b2-m is standardized with reference to highly purified proteins. The b2-m 8. Application of linear regression procedures for method
comparison studies. Part I. J Clin Chem Clin Biochem 1983;
a sensitive indicator of renal tubular disorders and has been used Material Required concentration of the Standard and Control is given on the label.
21:709-20.
to detected early nephotoxicity in patients treated with gentamicin Automatic analyzer. Prepare the following dilutions of the standards using saline solution:
9. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
and other nephrotoxic drugs. Besides renal insufficiency serum Saline solution. 1; 1/2; 1/4; 1/8; 1/16, saline. The standard dilutions are to be used
207 . 224.
level of b2-m was shown to be elevated in a variety of diseases Calibrator. for measurement within 4 hours. This curve is stored in memory
including carcinomas and lymphoid tumours and inflammatory and Controls. by the analyser and recalled for later use. Calibration curves are
autoimmune diseases such as Sjögren‘s syndrome and rheumatoid stable for up to 14 days, after which a new curve must be generated.
arthritis. Storage and Stability Additionally, recalibration must be performed whenever reagent lots
Detection of elevated serum b2-m has also been reported as a useful Reagents are ready to use. Shake the latex reagent gently before are changed. CATALOG NO QUANTITY
marker of acquired immune deficiency syndrome and in myeloma dispensing its content into the appropriate plastic vials. Reagents in
553 001 100 test
patients. This test provides the advantage of being the original bottle are stable to the expiration date indicated on the For quality control use Spectrum Control or other suitable control
economical, rapid, precise, accurate and suitable for the analysis of label when capped and stored at (2 - 8 ºC). Do not freeze. material. The control intervals and limits must be adapted to the
large series of serum and urine specimens. The b2- m buffer reagent should be clear and colourless. Any turbidity individual laboratory requirements. Values obtained should fall within
may be sign of deterioration and reagent should be discarded. The established limits. Each laboratory should
Test Principle b2- m latex reagent should have a white, turbid appearance free of establish corrective measures to be taken if values fall outside
The b2-m test is based upon the reactions between b2-m in granular particulate. Visible agglutination or precipitation may be a the limits. Control must be assayed and evaluated as for patient
the sample and latex-covalently bound rabbit antihuman b2-m sign of deterioration, and the reagent should be discarded. samples.
antibodies. b2-m values are determined turbidimetrically using fixed-
time measurement with sample blank correction. The relationship Specimen Collection and Preparation Calculation
between absorbance and b2-m concentration permits a multipoint Serum specimens should be collected by venipuncture following The turbidimetric analysers automatically calculate the b2- m
calibration with a measuring range between 0 and 12,0 mg/l. The good laboratory practices. Suitable assay specimens are human concentration of each sample.
measuring temperature is 37 ºC. serum samples, as fresh as possible (stored up to 7 days at 2 - 8 ºC) Conversion: mg/l = mg/ml.
The assay can be performed on all instruments allowing turbidimetric or deep-frozen. Any additional clotting or precipitation which occurs
measurements at 500 to 600 nm.. due to the freeze/thaw cycle should be removed by centrifugation Expected Values
prior to assay. Lipemic specimens, or turbid frozen specimens Values from 0,8 to 2,4 mg/l are within the normal range.
Reagents after thawing, must be clarified before the assay by high-speed This data must be interpreted as a guide. Each laboratory should
centrifugation (10 min at approx. 15.000 rpm). Heat-inactivation of
Buffer the serum samples can lead to a loss of the antigenic properties of
TRIS buffer containing detergents, polyethyleneglycol and < 0.1 % the b2- microglobulin and must therefore be avoided.
sodium azide as preservative.
System Parameter
Latex reagent Wavelength 600 nm
a suspension of latex microparticules covalently bound rabbit Optical path 1 cm
antihuman b2 -m antibodies in a glycine buffer, containing NaCL and Assay type Turbidimetric
bovine serum albumin. Preservative: Sodium azide < 0.1%. Temperature 37 ºC
180 181
b2 -MICROGLOBULIN SYMBOLS IN PRODUCT LABELLING Procedure References
Mono-Reagent Procedure EC REP Authorised Representative
Wavelength 600 nm 1. F. Ortega y col.. Influencia pronóstica de la b2-microglobulina
Temperature 37 ºC en el mieloma múltiple. Med. Clin. (Barc) 1992; 99: 645-648
Cuvette 1cm light path
REF Catalogue Number for use 2. Lifson A.R, Hessol N A, Buchbinder S P, O.Malley P M, Barnhart
REF: 554 002 100 test (R1 Buffer: 42.5 ml, R2 Latex: 8.5 ml) Measurement against distilled water blank. L, Segal M, Katz M H, Holmberg S D. Serum b2-microglobulin
Bring the reagents at 37ºC and pipette: and prediction of progression to AIDS in HIV infection. The
Lancet 1992; 339: 1436-40.
Intended Use Precautions and Warnings Calibrator Sample Blank 3. Weissel M, Scherak O, et al. Serum ß2-microglobulin and SLE.
In vitro diagnostic reagents for the quantitative determination of For in vitro diagnostic use only. Do not pipette by mouth. Reagents Arthritis Rheum 1976; 19:986.
Calibrator 3.5 mL ------ ------
b2 -Microglobulin (b2 -m) in serum by means of particle enhanced containing sodium azide must be handled with precaution. Sodium 4. Evrin P E, Paterson P A, Wide L, Berggard Y. Radioimmunoassay
Sample ------ 3.5 µl ------
turbidimetric immunoassay. azide can form explosive azides with lead and copper plumbing. of ƒÒ2- microglobulin in human biological fluids. Scand J clin
Distilled Water ------ ------ 3.5 µl Lab Invest 1971; 28: 439-43.
Background and reagents obtained from human blood should always be handled Work. Reagent 500 ml 500 ml 500 ml 5. Schardijn G, Statius van Eps L. b2-microglobulin: Its significance
b2-microglobulin (b2-m) , an 11,800-Da protein consisting of 100 with precaution using established good laboratory practices. in the evaluation of renal function. Kidney International 1987;
32: 635 41.
aminoacid residues, first isolated from urine by Berggard and Mix and measure absorbance immediately
Bearn, is located on the surface of lymphocytes and other nucleated Disposal of all waste material should be in accordance with local (A1) incubate 5 min (37ºC), after incubation read absorbance (A2). 6. Becker W. Die Standardddidierung immunchemischer
Plasmaprotein- Bestimmungen. Laboraatoriummblätter 1980;
cells and was identified to be the light chain of the class I major guidelines.
30: 25 . 32.
histocompatibility complex. The cell membrane turnover is the main Calculation
source of b2-m in serum, where its level rises when its production As with other diagnostic tests, results should be interpreted Plot the calibration curve and the sample concentration is obtained 7. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
207 . 224.
rate increases. Free b2-m is filtered by the glomerulus and considering all other test results and the clinical situation of the by interpolation the sample absorbance (A2-A1) in the calibration
subsequently reabsorbed almost completely in the proximal tubular patient. curve. 8. Application of linear regression procedures for method
cells, where it is catabolized. Increased urinary excretion of b2-m is If is an one point calibration:
21:709-20.
a sensitive indicator of renal tubular disorders and has been used Material Required
(A2-A1) sample - (A2-A1) blank 9. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
to detected early nephotoxicity in patients treated with gentamicin Spectrophotometric analyzer. x Calibrator concentration
207 . 224.
and other nephrotoxic drugs. Besides renal insufficiency serum Saline solution. (A2-A1) calibrator - (A2-A1) blank
level of b2-m was shown to be elevated in a variety of diseases Controls.
including carcinomas and lymphoid tumours and inflammatory and Linearity
autoimmune diseases such as Sjögren‘s syndrome and rheumatoid Storage and Stability Up to 20 mg/L. ORDERING INFORMATION
arthritis. Reagents in the original vial is stable to the expiration date on the vial CATALOG NO QUANTITY
Detection of elevated serum b2-m has also been reported as a useful label when capped and stored at (2 - 8 ºC). Immediately following the Calibration and Quality Control
marker of acquired immune deficiency syndrome and in myeloma completion of an assay run, the reagent vial should be capped until Calibrator 1 100 µl of Spectrum b2 -m Calibrator* 554 002 100 test
patients. This test provides the advantage of being next use in order to maximize curve stability. Once opened the Calibrator 2 100 µl of Calibrator 1 + 100 µl of Saline Solution
economical, rapid, precise, accurate and suitable for the analysis of reagent can be used within 1 month if stored tightly closed at Calibrator 3 100 µl of Calibrator 2 + 100 µl of Saline Solution
large series of serum specimens. (2 - 8 ºC) after use. Do not freeze reagents. Calibrator 4 100 µl of Calibrator 3 + 100 µl of Saline Solution
The b2 -Microglobulin latex reagent should have a white, turbid Calibrator 5 100 µl of Saline Solution
Test Principle appearance free of granular particulate. Visible agglutination or (*) See values on the label or on the insert. Multiply by the
This b2-m test is based upon the reactions between b2-m in the precipitation may be a sign of deterioration, and the reagent should appropriate factor.
sample and latex-covalently bound goat antihuman b2-m antibodies. be discarted.
b2-m values are determined photometrically. For quality control use Spectrum Control or other suitable control
The b2 -Microglobulin buffer reagent should be clear and colourless. material. The control intervals and limits must be adapted to the
Reagents Any turbidity may be sign of deterioration and reagent should be individual laboratory requirements. Values obtained should fall
discarted. within established limits. Each laboratory should establish corrective
Buffer measures to be taken if values fall outside the limits. Control must be
TRIS buffer, pH: 7,2, containing detergents, polyethyleneglycol and Specimen Collection and Preparation assayed and evaluated as for patient samples.
0,09 % sodium azide as preservative. Serum specimens should be collected by venipuncture following
good laboratory practices. Suitable assay specimens are human Expected Values
Latex reagenta serum samples, as fresh as possible (stored up to 7 days at 2 - 8 ºC) Values from 0,8 to 2,4 mg/L are within the normal range. Each
suspension of latex microparticules covalently bound goat antihuman or deep-frozen. Any additional clotting or precipitation which occurs laboratory should establish an expected range for the geographical
b2-m antibodies in a glycine buffer (0,1 M), containing NaCL (0,15 due to the freeze/thaw cycle should be removed by centrifugation area in which it is located.
M) and bovine serum albumin (0,5%). Preservative: Sodium azide prior to assay.
0,075%. Lipemic specimens, or turbid frozen specimens after thawing, must
be clarified before the assay by high-speed centrifugation (10 min at
Calibrator approx. 15.000 rpm).
Human - based reference fluid. Preservative: sodium azide, 0.075%. Heat-inactivation of the serum samples can lead to a loss of the
antigenic properties of the b2-m and must therefore be avoided.
All raw materials of human origin used in the manufacture of this
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 Reagent Preparation
and HCV with commercially available test methods. Working Reagent is prepared with 1 part of Latex Reagent and 9
However, this product should be handled as though capable of parts of Buffer Reagent. Prepare a fresh WR based on its workload.
transmitting infectious diseases Shake gently the reagents before pipetting.
182 183
C REACTIVE PROTEIN (CRP) SYMBOLS IN PRODUCT LABELLING
Quality Controls
Control sera are recommended to monitor the perfomance of
Immuno-Turbidimetry EC REP Authorised Representative
manual and automated assay procedures.
Each laboratory should establish its own Quality Control Scheme and
corrective actions if controls do not meet the acceptable tolerances.
REF 588 001 100 test Expected Values
R1 Buffer Reagent 2 x 20 ml 0 - 10 mg/L .
R2 Antiserum 1 X 4.2 ml
Reagent Preparation, Storage and Stability Each laboratory should establish an expected range for the
All reagents are supplied ready to use. geographical area in which it is located.
Reagents in the original vial are stable to the expiration date on the
Intended Use vial label when capped and stored at (2 - 8 ºC).
In vitro diagnostic reagents for the quantitative determination of
References
C Reactive Protein (CRP) in human serum by turbidimetric CRP Standard:
The Standard is stable to the expiration date on the vial label when 1. Ritchie, RF., J. Lab clin. Med. 70, 512 (1967)
immunoassay.
capped and stored at (2 - 8 ºC).
2. Pepys MB. et al., Ann. NY Acad. Sci, 389, 459 (1982)
Once opened the Standard is stable for 6 weeks if stored tightly
Background closed at 2 - 8 ºC after use. 3. Manack, J.R. and Richards, CB., J. Immunol. 20, 1019
C- reactive protein (CRP) is one of the acute phase proteins being For further storage at - 30 ºC divide Standard into aliquots. Stability
synthesised by hepatocytes. The serum concentration of CRP 3 months. once thawed never freeze again.
increases during acute stages of diverse diseases associated with
Specimen Collection and Preparation
inflammation and tissue injury. Elevated CRP has been demonstrated Fresh or deep frozen serum. CRP remain stable for 2 days at
in nearly all bacterial and fungal infections. In addition, it has been (2 - 8 ºC). If the test should be performed later, it is recommended to
freeze the serum. Avoid successive freezing and thawing. ORDERING INFORMATION
shown to be increased in other diseases as neoplasia, and rheumatic
diseases as well as in major surgery. The diagnosis usefulness of Discard haemolysed or contaminated samples. Heavily lipaemic CATALOG NO QUANTITY
CRP is based on the velocity and on the magnitude of its increase. sera and turbid frozen serum samples must be cleared with a
588 001 100 test
Serum concentrations are raised within hours of disease onset and delipidating agent. The cleared patient serum sample must be used
the increase can be as much 2000-fold. A rapid fall of CRP levels on the same day, as turbidity may reoccur.
indicates recovery.
Procedure
Test Principle 1. Bring the reagents and the photometer to room temperature
This CRP test is based upon the C reactive protein (CRP) 2. Assay conditions:
antigen-antibody reaction. Wavelength 340 nm
Temperature room temperature
Reagents Cuvette 1cm light path
R1 Buffer Reagent 3. Adjust the instrument to zero with distilled water .
Phosphate buffered saline(pH 7.43). 4. Pipette into a cuvette :
Polyethelene glycol (40 g/L). Standard Sample
Sodium azide (0.95 g/L). Reagent (R1) 400 ml 400 ml
Standard 25 ml ------
R2 Antiserum
Sample ------ 25 ml
Phosphate beffered saline(pH 7.43).
Polyclonal goat anti-human CRP (variable). Mix and record 1st reading (A1).
Materials required but not provided with the kit After addition of R2, incubate at room temperature and after 5
minutes record 2nd reading (A2)
1- Standard
Calculation
CRP concentration is stated on the vial label.
Generate a reference curve by successive 1 : 2 dilutions of calibrator
in saline (At Least 4 points are recommended). Use Saline as zero
2-Controls
point. Determine D absorbance of the sample and each calibrator
as following:
Precautions and Warnings D absorbance of sample = (A2 - A1) sample
For in vitro diagnostic use only. Do not pipette by mouth. Reagents D absorbance of each calibrator = (A2 - A1) for each standard
containing sodium azide must be handled with precaution. Sodium Plot the calibration curve and obtain the result.
azide can form explosive azides with lead and copper plumbing. Since
Sensitivity
absence of infectious agents cannot be proven, all specimens and
1 mg/L
reagents obtained from human blood should always be handled with
precaution using established good laboratory practices. Disposal of Linearity
all waste material should be in accordance with local guidelines. As Up to 220 mg/L.
with other diagnostic tests, results should be interpreted considering specimens showing higher concentration should be diluted 1+4
all other test results and the clinical situation of the patient. using physiological saline and repeat the assay (result×5).
184 185
C REACTIVE PROTEIN (CRP) SYMBOLS IN PRODUCT LABELLING Sensitivity
Turbi Latext EC REP Authorised Representative Temperature Limitation 2 mg/L
Linearity
REF: 560 001 100 test Up to 150 mg/L.
R1 Diluent 2 x 20 ml specimens showing higher concentration should be diluted 1+4
R2 Latex 1 X 10 ml using physiological saline and repeat the assay (result×5).
Quality Controls
Intended Use Storage and Stability Control sera are recommended to monitor the perfomance of manual
In vitro diagnostic reagents for the quantitative determination of Reagents in the original vial is stable to the expiration date on the vial and automated assay procedures.
C Reactive Protein (CRP) in human serum by means of particle- label when capped and stored at (2 - 8 ºC). Immediately following the Each laboratory should establish its own Quality Control Scheme and
enhanced turbidimetric immunoassay. completion of an assay run, the reagent vial should be capped until corrective actions if controls do not meet the acceptable tolerances.
next use in order to maximize stability. Do not freeze reagents.
Background The CRP latex reagent should have a white, turbid appearance free Expected Values
C- reactive protein (CRP) is one of the acute phase proteins being of granular particulate. Visible agglutination or precipitation may be a Values < 6 mg/L are within the normal range.
synthesised by hepatocytes. The serum concentration of CRP sign of deterioration, and the reagent should be discarted.
increases during acute stages of diverse diseases associated with The CRP diluent reagent should be clear and colourless. Any turbidity Each laboratory should establish an expected range for the
inflammation and tissue injury. Elevated CRP has been demonstrated may be sign of deterioration and reagent should be discarted. geographical area in which it is located.
in nearly all bacterial and fungal infections. In addition, it has been WR is stable for up to one month at 4ºC. It is recommended that
shown to be increased in other diseases as neoplasia, and rheumatic each Laboratory prepares a fresh Working Reagent based on its
References
diseases as well as in major surgery. The diagnosis usefulness of workload.
1. Hokama Y, Nakamura RM. C-Reactive protein: current status
CRP is based on the velocity and on the magnitude of its increase.
and future perspectives. J Clin Lab Anal 1987; 1: 15-27
Serum concentrations are raised within hours of disease onset and Reagent Preparation & Stability
the increase can be as much 2000-fold. A rapid fall of CRP levels Working Reagent is prepared with 1 part of Latex Reagent and 4 2. Hessian PA, Palmer DG. The presence and possible
significance of C-Reactive protein in rheumatoid inflammation.
indicates recovery. parts of Buffer Reagent . Prepare a fresh WR based on its workload. J Rheumatol 1985 1985; 12:871-5.
Shake gently the reagents before pipetting.
3. Okamura JM, Miyagi JM, Terada K, Hokama Y. Potential
Test Principle e.g. 400 ml Buffer Reagent + 100 ml Latex Reagent.
clinical
This CRP test is based upon the reaction between C reactive protein stability : 1 month at at 2 - 8 ºC.
4. applications of C-reactive protein. J Clin Lab Anal 1990;
(CRP) and latex covalently bound antibodies against human CRP. CRP Calibrator: Reconstitute with 1 ml distelled water. mix gently
4:231-5.
CRP values are determined photometrically. and incubate at room temperature for 10 minutes before use.
Stability: 1 month at at 2 - 8 ºC or 3 months at at -20 ºC 5. Müller M, Mierau R, Wohltmann D. Interference of IgM
rheumatoid factor with nephelometric C-reactive protein
Reagents determinations. J Immunol Methods 1985; 80: 77-90.
R1 Diluent 6. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
Fresh or deep frozen serum. CRP remain stable for 8 days at (2 -
TRIS buffer 20 mmol/L, pH 8.2, and 0.95 g/L sodium azide. Edition, AACC Press, 2000.
8 ºC). If the test should be performed later, it is recommended to
freeze the serum. Avoid successive freezing and thawing. 7. Passing H, Bablok W. A new biometrical procedure for testing
R2 Latex reagent the equality of measurements from two analytical methods.
Latex particles coated with goat IgG anti-human CRP, pH 7.3 and Discard haemolysed or contaminated samples. Heavily lipaemic
Sodium azide 0.95 g/L. sera and turbid frozen serum samples must be cleared with a 8. Application of linear regression procedures for method
delipidating agent. The cleared patient serum sample must be used
Calibrator 21:709-20.
on the same day, as turbidity may reoccur.
CRP concentration is stated on the vial label. 9. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
207 – 224
All raw materials of human origin used in the manufacture of this Procedure
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 1. Bring the reagents and the photometer to 37ºC
and HCV with commercially available test methods. However, 2. Assay conditions:
this product should be handled as though capable of transmitting Wavelength 540 nm (530 -550 nm)
infectious diseases Temperature 37ºC
Precautions and Warnings 3. Adjust the instrument to zero with distilled water . CATALOG NO QUANTITY
For in vitro diagnostic use only. Do not pipette by mouth. Reagents 4. Pipette into a cuvette
560 001 100 test
containing sodium azide must be handled with precaution. Sodium Working Reagent 500 ml
azide can form explosive azides with lead and copper plumbing. Since Calibrator or Sample 5 ml
absence of infectious agents cannot be proven, all specimens and 5.Mix and read absorbance immediately (A1) and after 2
reagents obtained from human blood should always be handled with min (A2) of the sample addition.
precaution using established good laboratory practices. Disposal of
all waste material should be in accordance with local guidelines. As Calculation
with other diagnostic tests, results should be interpreted considering
(A2-A1) sample
all other test results and the clinical situation of the patient. x Calibrator concentration = mg/L CRP
(A2-A1) calibrator
186 187
Ceruloplasmin SYMBOLS IN PRODUCT LABELLING CRP Standard Super High SYMBOLS IN PRODUCT LABELLING
Immuno-Turbidimetry EC REP Authorised Representative

EC REP Authorised Representative
REF Catalogue Number for use REF 347 000 1 x 0.5 ml REF Catalogue Number for use
REF: 592 001 50 test Consult instructions for use Manufactured by REF 347 001 1 x 1 ml Consult instructions for use Manufactured by
R1 Buffer Reagent 1 x 20 ml
R2 Antiserum 1 X 3 ml
Standard Sample
Reagent (R1) 400 ml 400 ml
Intended Use
Spectrum Diagnostics Ceruloplasmin reagent is intended for Standard 5 ml ------
Specifications
Quantitative turbidimetric determination of Ceruloplasmin in human Sample ------ 5 ml
serum .
Mix, incubate for 2 minutes and record 1st reading (A1).
Lot number : CRPS 3014
Background Reagent (R2) 60 ml 60 ml
Ceruloplasmin (CER) is a glycoprotein which is synthesized mostly Expiration Date : December 2015
in the liver. It is one of the acute phase protein in inflammation
After addition of R2, incubate at 37ºC and after 5 minutes record 2nd
and it is the most important carrier of copper (Cu) in plasma. The Source : Human plasma and pleural fluid
Ceruloplasmin molecule binds 6-8 Cu atoms. Ceruloplasmin has reading (A2)
antioxidative effect. The most important physiological functions Adaptation sheets for several automatic analyzers are available Storage: 2-8ºC Do Not Freeze
of Ceruloplasmin are the regulation of transport, availability, and upon request.
redox potential of iron (Fe) as a result of its ferroxidase activity; Stability after opening the vial : 6 weeks
the antioxidative effect of lipids in the cell membrane, due to the
Calculation
prevention of metal ion-catalyzed oxidation; and the transport of
Generate a reference curve by successive 1 : 2 dilutions of Standard
copper.
Assay Principle as following:
This Ceruloplasmin test is based upon the Ceruloplasmin
antigen-antibody reaction. D absorbance of sample = (A2 - A1) sample
D absorbance of each Standard = (A2 - A1) for each Standard
Reagent Plot the calibration curve and obtain the result.
R1 Buffer Reagent
Phosphate buffered saline(pH 7.43). Expected Values
Polyethelene glycol (40 g/L). Normal values are between 22 - 61 mg/dL.
Sodium azide (0.95 g/L). Each laboratory should establish its own reference range.
R2 Antiserum
Sensitivity
Polyclonal goat anti-human Ceruloplasmin (variable).
assay is 4 mg/dL.
Sodium azide (0.95 g/L).
Linearity
Materials required but not provided with the kit
Up to 100 mg/dL.
1-Standard
specimens showing higher concentration should be diluted 1/5 using
Ceruloplasmin concentration is stated on the vial label.
physiological saline and repeat the assay.
2-Controls
Interfering Substances:
Haemoglobin up to 1000 mg/dL.
Bilirubin up to 20 mg/dL.
Spectrum Ceruloplasmin reagents are stable up to the expiry date
Triglycerides up to 2500 mg/dL.
labeled on the bottles when stored at 2 - 8ºC and contaminations are
prevented during their use.Once opened the standard is stable for 6
weeks if stored tightly closed at 2 - 8 ºC after use. Dynamic Range Target CRP mg/L
For further storage at - 30 ºC divide standard into alicots. Stability 3 4 - 100 mg/dL.
CRP Standard Super High
months. once thawed never freeze again.
Waste Disposal 226.6
Specimen Collection and Preparation This product is made to be used in professional laboratories.
Fresh serum . Stable 2 days at 2 - 8ºC or 3 months at -20ºC. Please consult local regulations for a correct waste disposal.
Samples with presence of fibrin should be centrifuged. S56: dispose of this material and its container at hazardous or
Do not use highly hemolized or lipemic samples.Once opened the special waste collection point.
standard is stable for 6 weeks if stored tightly closed at 2 - 8 ºC after S57: use appropriate container to avoid environmental
use. contamination.
S61: avoid release in environment. refer to special
Procedure
Wavelength 340 nm
Temperature 37ºC References
Cuvette 1cm light path Poulik, M. D. and Weiss, M. L., in F. W. Putman, Editor, “The Plasma
Zero adjustment distilled water Proteins, vol. 2 second edition, academic press, New York,
pp. 52 - 108.
Bring the reagents at 37ºC and pipette:
188 189
CRP Control SYMBOLS IN PRODUCT LABELLING
REF 343 001 1 x 1 ml
Intended Use
Accuracy control for the determination of C- Reactive protiens in serum by turbidimetry and nephelometry.
Composition
A dilution of humen plasma and pleural fluid containing high levels of CRP with phosphsate buffered saline.
Liquid stabilised.
Preservative: 0.095 g% sodium azide .
The expiry date of the product at +2 to +8 ºC is listed on the label.

After first opening the control can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use.
Do not freeze.
1. For “In Vitro Diagnostic Use”

2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin
(HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were
used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in ac-
cordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven .
3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes !
Assigned Value
LOT No.: CRPT 3014
CRP : 35.5 ( 30.2 - 40.8 ) mg/L.
190 191
CRP/hs(C- Reactive protein) SYMBOLS IN PRODUCT LABELLING Procedure
EC REP Authorised Representative Temperature Limitation Wavelength 550 nm
IVD For in-vitro diagnostic use Use by/Expiration Date ORDERING INFORMATION
Temperature 37ºC
LOT Batch Code/Lot number CAUTION. Consult instructions CATALOG NO QUANTITY
REF: 545 001 50 test Cuvette 1cm light path
R1 Buffer 1 x 20 ml Measurement against distilled water blank.
Consult instructions for use Manufactured by 545 001 50 test
R2 Latex 1 x 4 ml Bring the reagents at 37ºC and pipette:
Blank Calibrator Sample

Intended Use Material Required but not provided
In vitro diagnostic reagents for the quantitative determination of Spectrophotometric analyzer. Buffer R1 375 ml 375 ml 375 ml
C Reactive Protein (CRP) in human serum by immunoassay. Saline solution. Latex Reagent R2 75 ml 75 ml 75 ml
Controls. Mix, Incubate one minute.
Background Distilled Water 5 ml --- ---
C- reactive protein (CRP) is one of the acute phase proteins being Storage and Stability Calibrator --- 5 ml ---
synthesised by hepatocytes. The serum concentration of CRP Reagents in the original vial is stable to the expiration date on the
Sample --- --- 5 ml
increases during acute stages of diverse diseases associated with vial label when capped and stored at (2 - 8 ºC). Immediately following
inflammation and tissue injury. Elevated CRP has been demonstrated the completion of an assay run, the reagent vial should be capped Mix and measure absorbance immediately (A1) ,
in nearly all bacterial and fungal infections. In addition, it has been until next use in order to maximize curve stability. Once opened then incubate 5 min (37ºC), after incubation read absorbance (A2).
shown to be increased in other diseases as neoplasia, and rheumatic the reagent can be used within 1 month if stored tightly closed at
diseases as well as in major surgery. The diagnosis usefulness of (2 - 8 ºC) after use. Do not freeze reagents. Calculation
CRP is based on the velocity and on the magnitude of its increase. The CRP latex reagent should have a white, turbid appearance free Plot the calibration curve and the sample concentration is obtained
by interpolation the sample absorbance (A2-A1) in the calibration
Serum concentrations are raised within hours of disease onset and of granular particulate. Visible agglutination or precipitation may be a
curve. If is an one point calibration:
the increase can be as much 2000-fold. A rapid fall of CRP levels sign of deterioration, and the reagent should be discarted.
indicates recovery. The CRP buffer reagent should be clear and colourless. Any turbidity
(A2-A1) sample - (A2-A1) blank
may be sign of deterioration and reagent should be discarted. x Calibrator concentration
Test Principle WR is stable for up to one month at 4ºC. It is recommended that (A2-A1) calibrator - (A2-A1) blank
This CRP test is based upon the reactions between C reactive each Laboratory prepares a fresh Working Reagent based on its
protein (CRP) and latexcovalently bound antibodies against human workload. Linearity
CRP. Up to 50 mg/L.
CRP values are determined turbidemetrically. Reagent Preparation
All reagent are supplied ready to use. Expected Values
Reagents Reagents in the original vial are stable to the expiration date on the Values < 6 - 8 mg/L are within the normal range.
Buffer R1 vial lable when capped and stored at (2 - 8 ºC).
Each laboratory should establish an expected range for the
TRIS buffer, pH: 6.5, and 0.09 % sodium azide as preservative. geographical area in which it is located.
Latex reagent R2 Fresh or deep frozen serum. CRP remain stable for 8 days at
References
Polystyrene particles (0.5%) coated with goat antibodies anti- (2 - 8 ºC). If the test should be performed later, it is recommended to
1. Hokama Y, Nakamura RM. C-Reactive protein: current status
human- CRP serum in aglycine buffer (0.1 M, pH: 8.2), containing freeze the serum. Avoid successive freezing and thawing.
and future perspectives. J Clin Lab Anal 1987; 1: 15-27
NaCL (0.15 M) and bovine serum albumin (0.5%). Preservative: Discard haemolysed or contaminated samples. Heavily lipaemic
Sodium azide 0.075% sera and turbid frozen serum samples must be cleared with a 2. Hessian PA, Palmer DG. The presence and possible
significance of C-Reactive protein in rheumatoid inflammation.
delipidating agent. Delipidation of do not affect the results of CRP in J Rheumatol 1985 1985; 12:871-5.
Calibrator serum samples. The cleared patient serum sample must be used on
3. Okamura JM, Miyagi JM, Terada K, Hokama Y. Potential
Human - based reference fluid. Preservative: sodium azide, 0.075% the same day, as turbidity may reoccur.
clinical applications of C-reactive protein. J Clin Lab Anal
Stability 4 weeks after opening. 1990; 4:231-5.
All raw materials of human origin used in the manufacture of this Calibration curve
4. Müller M, Mierau R, Wohltmann D. Interference of IgM
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 Calibrator 1 100 ml of Spectrum CRP Calibrator* rheumatoid factor with nephelometric C-reactive protein
and HCV with commercially available test methods. However, Calibrator 2 100 ml of Calibrator 1 + 100 ml of Saline Solution determinations. J Immunol Methods 1985; 80: 77-90.
this product should be handled as though capable of transmitting Calibrator 3 100 ml of Calibrator 2 + 100 ml of Saline Solution
Calibrator 4 100 ml of Calibrator 3 + 100 ml of Saline Solution 5. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
infectious diseases Calibrator 5 100 ml of Saline Solution Edition, AACC Press, 2000.
(*) See values on the label . Multiply by the appropriate factor.
6. Passing H, Bablok W. A new biometrical procedure for testing
Precautions and Warnings the equality of measurements from two analytical methods.
For in vitro diagnostic use only. Do not pipette by mouth. Reagents For quality control use Spectrum Control or other suitable control
7. Application of linear regression procedures for method
containing sodium azide must be handled with precaution. Sodium material. The control intervals and limits must be adapted to the comparison studies. Part I. J Clin Chem Clin Biochem 1983;
azide can form explosive azides with lead and copper plumbing. Since individual laboratory requirements. Values obtained should fall 21:709-20.
absence of infectious agents cannot be proven, all specimens and within established limits. Each laboratory should establish corrective
8. Sonderdruck aus DG Klinische Chemie Mitteilungen
reagents obtained from human blood should always be handled with measures to be taken if values fall outside the limits. Control must be 1995; 26: 207 – 224
precaution using established good laboratory practices. Disposal of assayed and evaluated as for patient samples.
all waste material should be in accordance with local guidelines. As
all other test results and the clinical situation of the patient.
192 193
CYSTATIN-C SYMBOLS IN PRODUCT LABELLING Calculation Dynamic Range
EC REP Authorised Representative Temperature Limitation (using calibration curve) 0 - 10 mg/L.
Immunoturbidimetry IVD For in-vitro diagnostic use Use by/Expiration Date
Determine D absorbance of the sample and each calibrator as Waste Disposal
following: This product is made to be used in professional laboratories.
REF: 570 002 100 test D absorbance of sample = (A2 - A1) sample Please consult local regulations for a correct waste disposal.
D absorbance of each calibrator = (A2 - A1) for each calibrator S56: dispose of this material and its container at hazardous or
Plot the calibration curve and obtain the result. special waste collection point.
Intended Use S57: use appropriate container to avoid environmental
Spectrum Diagnostics Cystatin-C reagent is intended for Quantitative Immediately following the completion of an assay run, the reagent (using single point) contamination.
turbidimetric determination of Cystatin-C in human serum. vials should be capped until next use in order to maximize curve Cystatin C concentration can be determined using single point S61: avoid release in environment. refer to special instructions/
stability. calibration (Calibrator No.2). safety data sheets.
Background
(A2-A1) sample
Cystatin C is a nonglycosilated 13-kDa basic protein belonging to Once opened the reagent can be used within 1 month if stored tightly x Calibrator concentration = mg/L Cystatin C
References
the cystatin super-family of cysteine proteinase inhibitors. Cystatin closed at 2 - 8 ºC after use. (A2-A1) calibrator
1. A V Lewis, T J James, J B J McGuire and R P Taylor. Improved
C is produced by virtually all nucleated cells, and is present in
immunoturbidimetric assay for cystatin C. Ann Clin Biochem
all investigated body fluids. The production rate is constant and Deterioration - If the sample result is abnormal value, we recommend repeating 2001; 38: 111 – 114.
unaffected by inflammatory processes, sex, age, diet and nutritional Do not use the Spectrum Cystatin C reagents if presence of particles the test using calibrator No.6 for more precission.
2. Mutsumi Tanaka, Kenje Matsuo, Masayasu Enomoto and
status. In the normal kidney, cystatin C is freely filtrated through the and turbidity. Koji Mizuno. A Sol particle homogeneus immunoassay for
glomerular membrane of the nephron and then nearly completely Do not freeze; frozen Antibody or diluent could change the Note: Adaptation sheets for several auto-analyzers are available measuring serum cystatin C. Cli. Biochem. 37 (2004) 27 – 35.
reabsorbed and degradated by the proximal tubular cells. Therefore, functionality of the test. upon request.
3. Davis Massey. Commentary: Clinical Diagnostic Use of
the plasma concentration of cystatin C is almost exclusively The Cystatin C latex reagent should have a white, turbid appearance Cystatin C. Journal of Clinical Laboratory Analysis 18:50 – 60
determined by the GFR (glomerular filtration rate), making cystatin free of granular particulate. Visible agglutination or precipitation may - The Turbidimetric analyzers automatically calculate the Cystatin C (2004).
C an excellent indicator of GFR. At the same time cystatin C is be a sign of deterioration, and the reagent should be discarded. concentration of each sample. 4. Michael G. Shlipak and al. Cystatin C and the risk of Death
becoming acknowledged as a marker of elevated risk of death from Conversion: mg/L = mg/ml. and cardiovascular events among elderly persons. NEJM 2005
cardiovascular complications – myocardial infarction and stroke. Precautions volume 352:2049-2060.
For in vitro diagnostic use only. Do not pipette by mouth. Reagents Calibration and Quality Control 5. David J Newman. Cystatin C. Ann Clin Biochem 2002; 39: 89
Assay Principle containing sodium azide must be handled with precaution. Sodium – 104.
This Cystatin C test is based upon the reactions between Cystatin azide can form explosive azides with lead and copper plumbing. The calibration curve is stored in memory by the analyzer and 6. Li Hai Xia, Xu Guo Bing and Xia Tie An. Serum Cystatin C
C and latex-covalently bound antibodies against human Cystatin Since absence of infectious agents cannot be proven, all specimens recalled for later use. assay for the detection of early renal impairment in diabetic
C. Cystatin C values are determined turbidimetrically using fixed- and reagents obtained from human blood should always be handled patients. JCLA 2004; 18:31-35.
time measurement with sample blank correction. The relationship with precaution using established good laboratory practices. Calibration curves are stable for up to 14 days, after which a 7. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
between absorbance and concentration permits a multipoint new curve must be generated. Additionally, recalibration must be Edition, AACC Press, 2000.
calibration with a measuring range between 0 and 10 mg/L. The Specimen Collection and Preparation performed whenever reagent lots are changed.
measuring temperature is 37 ºC. The assay can be performed on all Fresh or deep frozen serum. Cystatin C remains stable for 12 days ORDERING INFORMATION
instruments allowing turbidimetric measurements at 500 to 600 nm. at 2 - 8 ºC. If the test should be performed later, it is recommended For quality control use Spectrum-Diagnostics Control or other CATALOG NO QUANTITY
to freeze the serum. Avoid successive freezing and thawing. Discard suitable control material. The control intervals and limits must be
Reagent haemolysed or contaminated samples. adapted to the individual laboratory requirements. Values obtained 570 002 100 test
should fall within established limits. Each laboratory should establish
Buffer (R1) Procedure corrective measures to be taken if values fall outside the limits.
30 mL of TRIS buffer, pH: 7,2, containing detergents, Wavelength 550 nm Control must be assayed and evaluated as for patient samples.
polyethyleneglycol Temperature 37 ºC
Sodium azide 0.9 % Cuvette 1cm light path Expected Values
Zero adjustment distilled water Normal values are between 0.59 - 1.03 mg/L.
Latex Reagent (R2) Each laboratory should establish its own reference range.
5.1 ml of Polystyrene particles (0.5%) coated with antibodies Bring the reagents at 37ºC and pipette:
anti-human-Cystatin C serum in a glycine buffer (0.1 M, pH: 8.2), Sensitivity
containing NaCL (0.15 M) and bovine serum albumin (0.5%). Calibrator Sample When run as recommended, the minimum detection limit of the
Sodium azide 0.075 % Reagent (R1) 250 ml 250 ml assay is 0.05 mg/L.
Cystatin C Calibrators Linearity
Mix and incubate for 1 minute.
6 x 1 ml calibrators with different concentrations: Up to 10 mg/L.
Actual values are printed on the labels. Calibrator 3 mL ------ specimens showing higher concentration should be diluted using
Sample ------ 3 µl physiological saline and repeat the assay. Multiply the result by the
Reagent Preparation, Storage and Stability appropriate factor.
Spectrum Cystatin C reagents are stable up to the expiry date After addition of sample or calibrator record 1st reading (A1)
o
labeled on the bottles when stored at 2 - 8 C and contaminations immediately. Interfering Substances:
are prevented during their use. Incubate at 37ºC and after 5 minutes record 2nd reading (A2) Haemoglobin up to 5 g/L.
194 195
FERRITIN SYMBOLS IN PRODUCT LABELLING specimens showing higher concentration should be diluted 1+4
Calibration and Calibration curve
Use Ferritin calibrator.
using physiological saline and repeat the assay (result×5).
IVD Expected Values
The sensitivity of the assay and target value of calibrator have The determination of reference ranges for ferritin concentrations of
REF: 562 001 been statndardized against the 3rd international standard of ferritin clinically healthy individuals is very difficult. Ferritin concentrations
R1 Diluent 2 X 20 ml (94/572, 2008 WHO). Recalibrate when control results are out of are age- and sex- dependent and exhibit a wide range of distribution.
R2 Latex 1 X 10 ml specified tolerence, when using different lot of reagent and when
instrument is adjusted. Men 30 - 220 mg/L
Women 20 - 110 mg/LL
Intended Use Precautions and Warnings Calibration curve
In vitro diagnostic reagents for the quantitative determination of For in vitro diagnostic use only. Do not pipette by mouth. Reagents Calibrator dilution 1 2 3 4 These data are to be interpreted as a guide. Each laboratory should
containing sodium azide must be handled with precaution. Sodium
Ferritin in human serum by means of particle-enhanced turbidimetric Calibrator (ml) --- 25 50 100 establish its own reference intervals.
azide can form explosive azides with lead and copper plumbing.
immunoassay. Since absence of infectious agents cannot be proven, all specimens Na Cl 9 g/L (ml) 100 75 50 ---
and reagents obtained from human blood should always be handled
Factor 0 0.25 0.5 1.0 References
Background with precaution using established good laboratory practices.
Concentration (for example: 0 157 314 628 1. Wick M, Pinnggera W, Lehmann P. Ferritin in iron metabolism.
Ferritin is a macromolecule with a molecular weight of at least 440
Disposal of all waste material should be in accordance with local the undiluted C = 628 mg/L ) Diagnosis of anemias. 2nd ed. Springer-Verlag. Wien 1994.
kD and is formed of apoferritin and an iron core of about 2500 Fe+3 guidelines.
ions. It has been found a direct correlation between the plasma 2. Miles LEM, et al. Measurement of serum ferritin by a 2-site
As with other diagnostic tests, results should be interpreted immunoradiometric assay. Anal Biochem 1974; 61:209-224
ferritin concentration and the quantity of available iron stored in the Quality Control
considering all other test results and the clinical situation of the 3. Milmann N, Sondergaard M, Sorensen CM. Iron stores
body so that its determination is used for diagnosis and monitoring of Control sera are recommended to monitor the perfomance of manual
patient. in female blood donors evaluated by serum ferritin. Blut
iron deficiency and iron overload. Additional parameters (transferrin, and automated assay procedures. Each laboratory should establish
1985;51:337-345.
transferrin saturation, and haematological investigations) could its own Quality Control Scheme and corrective actions if controls do
4. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
be required for the diagnosis of disturbances of distribution. In a Storage and Stability not meet the acceptable tolerances.
Edition, AACC Press, 2000.
comparison of the various parameters available for the determination Reagents in the original vial is stable to the expiration date on the vial
of the body.s iron stores, plasma ferritin was the most efficient label when capped and stored at (2 - 8 ºC). Immediately following the Procedure 5. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
207 – 224
parameter, demonstrating a sensitivity of 80 %, and a specificity of completion of an assay run, the reagent vial should be capped until 1. Bring the reagents and the photometer to 37ºC
96 %. The serum concentrations of ferritin are found to be elevated next use in order to maximize curve stability. Do not freeze reagents. 2. Assay conditions:
in patients with infections, inflammation or in hepatic or chronic The Ferritin latex reagent should have a white, turbid appearance Wavelength 540 nm (530 -550 nm)
renal diseases. The determination of ferritin is particularly useful in free of granular particulate. Visible agglutination or precipitation may Temperature 37ºC ORDERING INFORMATION
the diagnosis of iron therapy, for the determination of iron reserves in be a sign of deterioration, and the reagent should be discarted. Cuvette 1cm light path CATALOG NO QUANTITY
high-risk groups, and in the differential diagnosis of anaemia. 3. Adjust the instrument to zero with distilled water .
The Ferritin diluent reagent should be clear and colourless. 4. Pipette into a cuvette : 562 001 100 test
Test Principle Any turbidity may be sign of deterioration and reagent should be Diluent (R1) 400 ml
This Ferritin test is based upon the reactions between Ferritin in discarted. Latex (R2) 100 ml
the sample and latexcovalently bound rabbit antihuman Ferritin Calibrator or Sample 45 ml
antibodies. Ferritin values are determined photometrically. Specimen Collection and Preparation 5.Mix and read absorbance immediately (A1) and after 5 min (A2) of
Specimens should be collected by venipuncture following good the sample addition.
Reagents laboratory practices. Suitable assay specimens are human serum
R1 Diluent samples, as fresh as possible (stored up to 7 days at 2 - 8 ºC) or Calculation
Trisbuffer 20mmol/L,pH8.2.Sodium azide0.95 g/L. deep-frozen. Any additional clotting or precipitation, which occurs Calculate the absorbance difference (A2-A1) of each point of the
calibration curve and plot the values obtained against the ferritin
due to the freeze/thaw cycle, should be removed by centrifugation
concentration of calibrator dilution. Ferritin concentration in the
R2 Latex reagent prior to assay. sample is calculated by interpolation of its (A2-A1) in the calibration
Latex particles coated with streptolysin O,pH 8.2 Very lipemic specimens, or turbid frozen specimens after thawing, curve.
Sodium azide 0.95 g/L. must be clarified before the assay by high-speed centrifugation (15
min at approx. 15.000 rpm). Sensitivity
Calibrator Up to 5.04 mg/L.
Human serum.Ferritin concentration is stated on the vial label. Reagent Preparation and Stability
Spectrum Ferritin reagents (R1 & R2) are supplied ready-to-use and Linearity
All raw materials of human origin used in the manufacture of this stable up to the expiry date labeled on the bottles when properly Up to 600 mg/L.
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 stored refrigerated at 2 – 8 oC. Once opened.
and HCV with commercially available test methods. However,
this product should be handled as though capable of transmitting Ferritin Calibrator: Reconstitute with 3 ml distelled water. mix
infectious gently and incubate at room temperature for 10 minutes before use.
diseases
Stability: 1 month at at 2 - 8 ºC or 3 months at at -20 ºC.
196 197
FIBRINOGEN SYMBOLS IN PRODUCT LABELLING
Example :
Immuno - Turbidimetry EC REP Authorised Representative

REF: 590 001 50 test

Intended Use All reagents are supplied ready to use.
In vitro diagnostic reagents for the quantitative determination of Reagents in the original vial are stable to the expiration date on the
Fibrinogen in human plasma by turbidimetric immunoassay. vial label when capped and stored at (2 - 8 ºC).
Background Fibrinogen Standard:

Minimizing blood loss is accomplished by three events.One is a Reconstitute with 0.5 ml distelled water. mix gently and incubate at
clumping of platelets in the blood at the site of injury.Another is a room temperature for 30 minutes before use.
vasoconstriction of the injured vessel to reduce the flow through Stability: 48 hours at 2 - 8 ºC or 2 weeks at at -20 ºC
the break.The third event is aggregation of a protein, fibrin, into a Sensitivity
clot – a stable three-dimensional lattice- that is strong enough to Note: Standard should be diluted 1 : 10 in saline before use. 4.5 mg/dL
seal the damaged vessel while repairs are being made. Clotting
occurs because a soluble blood plasma protein, fibrinogen, is Specimen Collection and Preparation Linearity
partially hydrolysed to form fibrin. Elevated levels of fibrinogen Fresh or deep frozen citrate plasma. fibrinogen remain stable Up to 523 mg/L.
specimens showing higher concentration should be diluted 1+4
in plasma are to be expected in inflammatory processes, after for 2 days at (2 - 8 ºC). If the test should be performed later, it is using physiological saline and repeat the assay (result×5).
major trauma or surgery and also occur with metastasing recommended to freeze the serum. Avoid successive freezing and
tumours.•Decreased levels of fibrinogen can occur in consumption thawing.Discard haemolysed or contaminated samples. Quality Controls
coagulopathies, e.g. disseminated intravascular coagulation (DIC), Control sera are recommended to monitor the perfomance of manual
Note: Sample should be diluted 1 : 10 in saline before use. and automated assay procedures.
primaryhyperfibrinolysis, hepatic insufficiency and genetic deficiency.
Each laboratory should establish its own Quality Control Scheme and
Epidemiological studies have shown that elevated plasma levels of corrective actions if controls do not meet the acceptable tolerances.
Procedure
fibrinogen are associated with an increased risk of arteriosclerosis. 1. Bring the reagents and the photometer to 37 ºC
2. Assay conditions: Expected Values
Test Principle Wavelength 340 nm 200 - 400 mg/dL .
Temperature 37ºC Each laboratory should establish an expected range for the
This Fibrinogen test is based upon the Fibrinogen antigen-antibody geographical area in which it is located.
reaction. 3. Adjust the instrument to zero with distilled water .
4. Samples, Controls and Standard should be diluted 1 : 10 in References
Reagents saline. 1. Dati. F. et al., Klin. Lab 39, 669 (199 3)
R1 Buffer Reagent 5. Pipette into a cuvette : 2. Ernst, E. und Resch, K. L., Ann. Intern. Med. 118, 956 (1993)
Phosphate buffered saline(pH 7.43). 3. Cremer, P. et al., Diagnose & Labor 42, 28 (1992 )
Enhancer. Standard Sample
Standard (diluted) 5 ml ------ ORDERING INFORMATION
R2 Antiserum
Sample (diluted) ------ 5 ml CATALOG NO QUANTITY
Polyclonal goat anti-human Fibrinogen (variable). Mix, incubate for 2 minutes and record 1st reading (A1). 590 001 50 test
Sodium azide (0.95 g/L). Reagent (R2) 50 mL 50 mL
After addition of R2, incubate and after 5 minutes record 2nd reading
(A2)
1- Standard
Fibrinogen concentration is stated on the vial label.
Calculation
2-Controls Generate a reference curve by successive 1 : 2 dilutions of Standard
in saline ( 6 Points ). Use Saline as zero point. Determine
D absorbance of the sample and each calibrator as following:
Precautions and Warnings D absorbance of sample = (A2 - A1) sample
For in vitro diagnostic use only. Do not pipette by mouth. Reagents D absorbance of each standard = (A2 - A1) for each Standard
containing sodium azide must be handled with precaution. Sodium Plot the calibration curve and obtain the result.
azide can form explosive azides with lead and copper plumbing. Since
absence of infectious agents cannot be proven, all specimens and
reagents obtained from human blood should always be handled with
precaution using established good laboratory practices. Disposal of
all waste material should be in accordance with local guidelines. As
all other test results and the clinical situation of the patient.
198 199
Fibrinogen Standard SYMBOLS IN PRODUCT LABELLING
REF 354001 1 X 0.5 ml LOT Batch Code/Lot number CAUTION. Consult instructions
Intended Use
Preparation of refrence curves for quantitaives immunochemical determination of Fibrinogen in cittrated plasma by turbidimetry and
nephelometry.
composition
A lyophilized preparation of stabilized, buffered human plasma containing less than 5% bovine plasma.
Preparation
1. Reconstitute the Fibrinogen standard with 0.5 ml of distilled or deionised water.
2. Restopper vial and allow to stand until fully hydrated (a minimum of 5 minutes).
3. Gently mix by rotation or inversion untill dissolution is complete. DO NOT SHAKE.
4. Allow to equilibrate for a total of 30 minutes befor use .

Stored at +2 to +8 c the Fibrinogen standard remains stable up to labelled expiry date .After reconstitution ,the Fibrinogen standard is stable
for 48 hours at +2 to +8 C when stored in the original vial in a sealed polypropylene container.

1. Fibrinogen standard is for “In Vitro Diagnosticc Use”
2. Fibrinogen standard is apotentially biohazardous material. Source materials rom which this product was derived were found negative for
HBsAg and for antibodies againist HCV,HIV-1 and HIV-2 by approved test methods.However,since no test method can offer complete
assurance that infectious agents are absent ,this product should be handled observing the same safety precautions employed when
handling any potentialy infectios material.
3. Fibrinogen Standard is Harmful . Harmful in contact with skin .Irritating to eye ,respiratory system and skin . In case of contact with
eyes,rinse immediately with plenty of water and seek medical advice.
200 201
HAPTOGLOBIN (HAP) SYMBOLS IN PRODUCT LABELLING Hemoglobin A1c SYMBOLS IN PRODUCT LABELLING
Immuno Turbidimetry EC REP Authorised Representative

Standard Set EC REP Authorised Representative
REF: 594 001 50 test
Intended Use
Intended Use Standard Sample Accuracy control in the normal range for quantitaive determination of HbA1c in human blood by turbidimetric imunoassay.
Spectrum Diagnostics Haptoglobin reagent is intended for Reagent (R1) 400 ml 400 ml
Quantitative turbidimetric determination of Haptoglobin in human composition
Dil.Standard 5 ml ------
serum . The HbA1c Standard set level 1 is a liquid buffer with preservative :0.095 % sodium azide .the HbA1c standard set level 2 , level 3 are
Dil. Sample ------ 5 ml
hemolysates prepared from packaged human erythrocytes, lyophilized and stabilised.
Background Mix, and record 1st reading (A1).
Acute phase protein.Transport molecule for haemoglobin. Reagent (R2) 60 mL 60 mL Preparation

Increased levels of HAP are reported in acute inflamation, 1. HbA1c Standard Set Level 1 ,ready for use.
After addition of R2, incubate at room tepmperature and after 5
collagenoses, coronary disorders, Hodgkin’s disease, nephrotic minutes record 2nd reading (A2) 2. HbA1c Standard Set Level 2,Level 3,accrately add 0.5 ml of distilled water to dissolve the contents,Gently mix for 10 minutes ,or untill
syndrom and tuberculosis. Decreased levels of HAP are found in all material has dissolved.
haemolytic anemia, liver disease, congenital deficiencies and acute Calculation
malaria. Generate a reference curve by successive 1 : 2 dilutions of diluted Storage and Stability
Standard in saline ( 4 Points ). Use Saline as zero point. Determine
D absorbance of the sample and each standard as following: The expiry date of the product at +2 to +8 c is listed on the label.
Assay Principle D absorbance of sample = (A2 - A1) sample The reconstituted standard set is stable for 15 days if stored tightly closed at +2 to +8 c
This HAP test is based upon the haptoglobin antigen-antibody D absorbance of each standard = (A2 - A1) for each Standard
reaction. Plot the calibration curve and obtain the result. Procedure
The lyophylized HbA1c standard set will produce a calibration curve that will be stable for at least 1 week on most analyzers.The standards
Expected Values
Reagents Normal values are between 32 - 205 mg/dL. should be treated in the same manner as patient sampless regarding the hemolysate procedure.Fllow the directions that accompany the
R1 Buffer Reagent Each laboratory should establish its own reference range. instruments and reagent kit used in the assay for specific instruments calibration procedures.
Phosphate buffered saline(pH 7.43).
Polyethelene glycol (60 g/L).
Sodium azide (0.95 g/L). Sensitivity Precautions and Warnings
When run as recommended, the minimum detection limit of the 1. For “In Vitro Diagnosticc Use”
R2 Antiserum assay is 1 mg/dL.
2. All blood doner units comparising the source material used in the manufacture of this product have been tested by approved FDA
Linearity methods and found to be non-reactive for HBsAg,HCV and anti-HIV 1 & 2. Only nevertheless every product obtained from human
Polyclonal goat anti-human Haptoglobin (variable).
Sodium azide (0.95 g/L). Up to 500 mg/dL. body fluides should be handled with appropriate care in accordance with recommended procedures for biohazardous materials since
specimens showing higher concentration should be diluted 1/5 using absence of infectious agents can never be proven .
Materials required but not provided with the kit physiological saline and repeat the assay.
1- Standard Interfering Substances: STS 1 STS 2 STS 3

Haptoglobin concentration is stated on the vial label. Haemoglobin up to 1000 mg/dL. % % %
2-Controls Triglycerides up to 2500 mg/dL.
HbA1c 0.0 5.2 14.8
Reagent Preparation, Storage and Stability Dynamic Range

Spectrum Haptoglobin reagents are stable up to the expiry date 1 - 500 mg/dL
Values based on reference material from NGSP/DCCT
labeled on the bottles when stored at 2 - 8oC and contaminations
are prevented during their use.Once opened the standard is stable Waste Disposal
for 6 weeks if stored tightly closed at 2 - 8 ºC after use. This product is made to be used in professional laboratories.
For further storage at - 30 ºC divide standard into aliquots. Stability Please consult local regulations for a correct waste disposal.
3 months. once thawed never freeze again. S56: dispose of this material and its container at hazardous or
Specimen Collection and Preparation S57: use appropriate container to avoid environmental
Fresh serum. Stable 2 days at 2 - 8 ºC or 3 months at -20 ºC. contamination.
Samples with presence of fibrin should be centrifuged. S61: avoid release in environment. refer to special
Do not use highly hemolized or lipemic samples.
Note: Sample should be diluted 1 : 10 in saline before use.
References
1. Dati, F. et al., Lab. Med. 13, 87 (1989)
Procedure
Wavelength 340 nm
Temperature room tepmperature ORDERING INFORMATION
CATALOG NO QUANTITY
Zero adjustment distilled water
Bring the reagents at room tepmperature
592 001 100 test
Samples, standard and controls should be diluted 1 : 10 in saline
before use.
202 203
Hemoglobin A1c (HbA1c) SYMBOLS IN PRODUCT LABELLING
Waste Disposal
Immuno - Turbidimetry EC REP Authorised Representative
LOT Batch Code/Lot number CAUTION. Consult instructions special waste collection point.
REF Catalogue Number for use S57: use appropriate container to avoid environmental
contamination.
REF: 602 001 50 test S61: avoid release in environment. refer to special
Reagent1 1 x 20 ml instructions/safety data sheets.
Reagent2 1 X 4 ml
References
1. Bates, H.M., Lab. Mang., Vol 16 (Jan. 1978)

Intended Use Hemolysate procedure 2. Gonen, B., and Rubenstein, A.H., Diabetologia 15, 1 (1978).
Spectrum Diagnostics Hemoglobin A1c reagent is intended for To determine HbA1c, a hemolysate must be prepared for each 3. Trivelli, L.A., Ranney, H.M., and Lai, H.T., New eng. J. Med.
sample as follow:
Quantitative turbidimetric determination of HbA1c in human blood 284, 353 (1971).
1. Dispense 2 ml hemolysis reagent into a test tube.
2. Place 20 ml of well mixed whole EDTA blood (Samples,
Background Standards and Controls) into the test tube and mix.
The glycemic control in diabetes mellitus is mainly by the 3. Allow to rest 5 minutes or until complete lysis is evident. Stability
of the hemosylate: 72 hours at 2 - 8 ºC.
determination of glucose, but also through quantitative determination ORDERING INFORMATION
of hemoglobin A1c in human blood. HbA1c is an indication for the Procedure CATALOG NO QUANTITY
actual glucose levels over the preceding 3 months. It was shown Wavelength 650 nm
that HbA1c in diabetic subjects can be elevated 2-3 fold over normal Temperature 37 ºC 602 001 50 test
and on other hand approaches normal values when they are under Cuvette 1cm light path
Zero adjustment distilled water
metabolic control.
Solve and lyse standard/control
Assay Principle
This method utilizes the interaction of antigen and antibody to Standard Sample
determine th HbA1c in whole EDTA blood. HbA1c in test samples Reagent (R1) 375 ml 375 ml
is absorbed onto the surface of latex particles, whiche react with
Anti-HbA1c (antigen-antibody reaction)and gives agglutination. The
Sample ------ 5 ml
amount of agglutination is measured as absorbance. The HbA1c
value is obtained from a calibration curve. Mix, and incubate for 2 minutes, then add
Reagent (R2) 75 mL 75 mL
Reagents
Reagent1 (R1) Mix and read absorbance (A1), incubate for 5 minutes and read
Latex. absorbance (A2)
Calculation
Reagent2 (R2) Generate a reference curve using HbA1c standard set. Determine
Anti-human hemoglobin A1c mouse monoclonal antibody. D absorbance of the sample and each standard as following:
Stabilizers. D absorbance of sample = (A2 - A1) sample
D absorbance of each standard = (A2 - A1) for each Standard
Materials required but not provided with the kit Plot the calibration curve and obtain the result.
1- Standard set Expected Values

HbA1c concentration is stated on the vials labels. Non-diabetics <6%
Theraputic diabetics <7%
2-Controls Each laboratory should establish its own reference range.
Reagent Preparation, Storage and Stability Linearity

Spectrum HbA1c reagents are stable up to the expiry date labeled on Up to 15 %.
the bottles when stored at 2 - 8 ºC and contaminations are prevented specimens showing higher concentration should be diluted 1/5 using
during their use.Once opened the reagents are stable for 1 month if physiological saline and repeat the assay.
stored tightly closed at 2 - 8 ºC after use.
Dynamic Range
Specimen Collection and Preparation 0 - 15 %.
Fresh EDTA blood.
204 205
For quality control use Spectrum Control or other suitable control
IMMUNOGLOBULIN E (IgE) SYMBOLS IN PRODUCT LABELLING material. The control intervals and limits must be adapted to the References
EC REP Authorised Representative Temperature Limitation individual laboratory requirements. Values obtained should fall
1. Kjellman NIM, Johansson SGO, Roth A. Clinical Allergy 1976;
REF: 548 001 50 test IVD For in-vitro diagnostic use Use by/Expiration Date within established limits. Each laboratory should establish corrective
6:51-59
R1 Buffer 1 x16 ml LOT Batch Code/Lot number CAUTION. Consult instructions measures to be taken if values fall outside the limits. Control must be
R2 Latex 1 x 6 ml Catalogue Number for use
assayed and evaluated as for patient samples. 2. Debelic,M. Clinical Significance of total and specific IgE in
REF
Consult instructions for use Manufactured by bronchial asthma. Allergol Immunopathol 1976;4: 361-70.
Procedure
3. Grundbacher, F,J, Causes of variation in serum IgE levels, in
Wavelength 600 nm normal population. J All Clin Immunol. 1975;56:104-11.
Intended Use Precautions and Warnings Optical path 1 cm
4. Dati, F. Ringel, K. Refernce values for serum IgE in healthy non
In vitro diagnostic reagents for the quantitative determination of For in vitro diagnostic use only. Do not pipette by mouth. Reagents Assay type Turbidimetric atopic children and adults. Clin Chem. 1982; 28:1556.
Immunoglobulin E (IgE) in human serum by the Immunoturbidimetric containing sodium azide must be handled with precaution. Sodium Temperature 37 ºC
5. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
procedure. azide can form explosive azides with lead and copper plumbing. Since Incubation time 4 min.
207 .224
absence of infectious agents cannot be proven, all specimens and Measurement against reagent blank.
Background reagents obtained from human blood should always be handled with Bring the reagents at 37ºC and pipette:
The Immunoglobulin E (IgE) has a molecular weight of aprox. precaution using established good laboratory practices. Disposal of
190000 g/mol and is produced by the organism in small quantities. all waste material should be in accordance with local guidelines. As Blank Calibrator Sample
Allergic diseases are a sign of hypersensitivity of the body. The type with other diagnostic tests, results should be interpreted considering CATALOG NO QUANTITY
Buffer 300 µl 300 µl 300 µl
I hypersensitivity reaction, also called immediate hypersensitivity, is all other test results and the clinical situation of the patient.
Latex reagent R2 110 µl 110 µl 110 µl 548 001 50 test
IgE mediated and is characterised by an immediate reaction following
Mtix, Incubate one minute.
contact with the antigen. Antigens facilitating an IgE response include Material Required
components of grass pollen, components of food, parasites and Automatic analyzer. Distilled Water 20 µl ------ ------
secretions from insects. This antigen induces the mucosal-B-cells, Saline solution. Calibrator ------ 20 µL ------
in conjunction with T-helper cells, to produce specific IgE. The IgE Sample ------ ------ 20 µl
molecules bind via Fc receptors to mast cells, which thus becomes Storage and Stability
sensitized. The next time when the antigen comes into contact with The IgE reagents should be stored tightly capped at (2 - 8 ºC) when For Blank, Calibrator and Sample, Mix and measure First absorbance
after 2 seconds (A1), then incubate 4 min, after incubation read
the sensitised mast cells, the bound IgE antibodies become cross- not in use. Do not freeze. Reagents in the original vials are stable
Second absorbance (A2).
linked, leading to degranulation of the mast cells and release of to the expiration date on the vial label when capped and stored at
mediators (as Histamine). The mediators bring about clinical signs (2 - 8 ºC). Calibration and Quality Control
typical for allergy, such as rhinitis, urtecaria, asthma and eczema. Standardization: use Spectrum Calibrators. The method was
IgE is formed mainly in the lymph nodes and mucous membranes Immediately following the completion of an assay run, the reagent standardized against to IRP 75/502. For quality control use Spectrum
of the respiratory and gastrointestinal tracts. IgE molecules cannot vials should be capped until next use in order to maximize curve Control or other suitable control material. Each laboratory should
pass through the placental barrier and do not activate complement. stability. Once opened the reagent can be used within 1 month if establish corrective measures to be taken if values fall outside
IgE determinations are indicated in the diagnosis and monitoring of stored tightly closed at (2 - 8 ºC) after use. The IgE buffer reagent the limits. Control must be assayed and evaluated as for patient
allergic diseases. should be clear and colourless. Any turbidity may be sign of samples.
Elevated IgE levels also occur in parasitosis and immunodeficiency deterioration and reagent should be discarted.
syndromes, such as acquired T-cell deficiency or the Wiskott-Aldrich Calculation
syndrome. In infants and small children with recurrent respiratory The IgE latex reagent should have a white, turbid appearance free
tract diseases (bronchitis, pseudocroup attacks), the determination of granular particulate. Visible agglutination or precipitation may be a x Calibrator concentration
of IgE is of prognostic relevance, also in some mielomas of IgE type. sign of deterioration, and the reagent should be discarted. (A2-A1) calibrator - (A2-A1) blank
Test Principle Specimen Collection and Preparation Expected Values

The Spectrum IgE test is used for the quantitative in vitro determination Serum specimens should be collected by venipuncture following The serum IgE concentration in healthy, nonatopic test subjects is
of total immunoglobulin IgE in serum and plasma samples. Anti-IgE good laboratory practices. very age dependent.
antibodies covalently bound to latex particles react with the antigen Suitable assay specimens are human serum samples, as fresh
(IgE) in the sample to form an antigen-antibody reaction complex, as possible (stored up to 2 days at 2 - 8 ºC) or deep-frozen. Any Age IU/ml
which can be measured turbidimetrically after particle aggregation. additional clotting or precipitation which occurs due to the freeze/
thaw cycle should be removed by centrifugation prior to assay. New-borns < 1.5
Reagents Very Lipemic specimens, or turbid frozen specimens after thawing, Infants<1 year < 15
Buffer R1 must be clarified before the assay by high-speed centrifugation (15 Children (1-5 years of age) < 60
Phosphate buffer, pH:7,0 , containing protein stabilizers and < 0.1 %
min at approx. 15.000 rpm). Children (6-9 years of age) < 90
sodium azide as preservative. Free of polyethyleneglicol.
Heat inactivation of serum samples results in loss of IgE antigenicity Children (10-15 years of age) < 200
Latex reagent R2 and therefore must be avoided. Adults < 100
Suspension of latex microparticules covalently bound anti-IgE These data are to be interpreted as a guide. Each laboratory should
antibodies suspended in a neutral aqueous solution, with < 0.1 %
Calibrator 1 100 ml of Spectrum IgE Calibrator* establish its own reference intervals.
sodium azide as preservative.
Calibrator 2 100 ml of Calibrator 1 + 100 ml of Saline Solution
Calibrator Calibrator 3 100 ml of Calibrator 2 + 100 ml of Saline Solution
Human - based reference fluid. Preservative: sodium azide, 0.075% Calibrator 4 100 ml of Calibrator 3 + 100 ml of Saline Solution
Ready to use ; stablity 4 weeks after opening. Calibrator 5 100 ml of Saline Solution
Actual value is printed on the vial label. (*) See values on the label . Multiply by the appropriate factor.
206 207
Immunology Control High SYMBOLS IN PRODUCT LABELLING Immunology Control Low SYMBOLS IN PRODUCT LABELLING
REF: 608 001 1x1 ml IVD For in-vitro diagnostic use Use by/Expiration Date REF: 606 001 1x1 ml IVD For in-vitro diagnostic use Use by/Expiration Date
REF: 608 002 2x1 ml REF: 606 002 2x1 ml
Intended Use Intended Use

Accuracy control for the determination of protiens in serum by turbidimetry and nephelometry. Accuracy control for the determination of protiens in serum by turbidimetry and nephelometry.
composition composition
A dilution of humen plasma with phosphsate buffered saline. A dilution of humen plasma with phosphsate buffered saline.
Contains stabilizers and 0.095 g% sodium azide . Contains stabilizers and 0.095 g% sodium azide .
Storage and Stability Storage and Stability

The expiry date of the product at +2 to +8 ºC is listed on the label. The expiry date of the product at +2 to +8 ºC is listed on the label.
After first opening the control can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use. After first opening the control can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use.
Do not freeze. Do not freeze.
Precautions and Warnings Precautions and Warnings

1. For “In Vitro Diagnosticc Use” 1. For “In Vitro Diagnosticc Use”
2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin 2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin
(HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were (HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were
used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in ac- used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in ac-
cordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven . cordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven .
3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes ! 3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes !
Lot No.: PC2021 Lot No.: PC1017
Protein Protein Values(mg/dL) Protein Protein Values(mg/dL)

Target Range Target Range
Albumin 3708 3152 - 5264 Albumin 1864 1584 - 2144
A1-Acidglycoprotein 83.2 70.7 - 95.7 A1-Acidglycoprotein 40.9 34.8 - 47.0
A1-Antitrypsin 154 131 - 177 A1-Antitrypsin 81.9 69.6 - 94.2
A2-Macroglobulin 218 185 - 251 A2-Macroglobulin 116 99 -133
ASL(IU/ml) 159 135 - 185 ASL(IU/ml) 82.9 70.5 - 95.3
Antithrombin III 25.0 21.3 - 28.8 Antithrombin III 12.8 10.9 - 14.7
B2-Microglobulin(mg/L) 4.08 3.47- 4.69 B2-Microglobulin(mg/L) 2.10 1.79 - 2.42
Ceruloplasmin 27.6 23.5 - 31.7 Ceruloplasmin 17.4 14.8 - 20.0
Complement C3 151 128 - 174 Complement C3 86.5 73.5 - 99.5
ComplementC4 30.5 25.9 - 35.1 ComplementC4 16.1 13.7 -18.5
C1 esterase inhibitor 38.0 32.3 - 43.7 C1 esterase inhibitor 16.8 14.3 -19.3
CRP 4.31 3.66 - 4.96 CRP 2.10 1.79 - 2.42
Ferritin(ng/mL) 174 148 - 200 Ferritin(ng/mL) 90.3 76.8 - 104
Haptoglobin 141 120 - 162 Haptoglobin 75.8 64.4 - 87.2
IgA 266 226 - 306 IgA 149 127 - 171
IgG 1121 953 - 1289 IgG 645 548 - 742
IgM 152 129 - 175 IgM 86.7 73.7 - 99.7
Kappa Light Chain 297 252 - 342 Kappa Light Chain 161 137 - 185
Lambada Light Chain 163 139 - 187 Lambada Light Chain 90.4 76.8 - 104
Prealbumin 35.8 30.4 - 41.2 Prealbumin 16.6 14.1 - 19.1
RF(IU/mL) 59.4 50.5 - 68.3 RF(IU/mL) 34.8 29.6 - 40.0
Transferrin 236 201-271 Transferrin 128 109 - 147
Values based on a reference preparation (ERM-DA470K) from the International Federation of Clinical Chemistry (IFCC). Values based on a reference preparation (ERM-DA470K) from the International Federation of Clinical Chemistry (IFCC).
Values based on WHO standard material. Values based on WHO standard material.
208 209
Kappa light chain serum kit SYMBOLS IN PRODUCT LABELLING Kappa light chain urine kit SYMBOLS IN PRODUCT LABELLING
(KAP) EC REP Authorised Representative

(KAP) EC REP Authorised Representative
Quantitative determination of Kappa light chain (KAP) in REF Catalogue Number for use
human serum by turbidimetric immunoassay. Consult instructions for use Manufactured by human urine by turbidimetric immunoassay. Consult instructions for use Manufactured by
REF: 375 001 50 test REF: 378 001 50 test

R1 Buffer Reagent 1 x 18 ml Procedure R1 Buffer Reagent 1 x 13 ml Procedure
R2 Antiserum 1 X 4 ml 1. Bring the reagents to 37 oC R2 Antiserum 1 X 1.7 ml 1. Bring the reagents to 37 oC
2. Assay conditions: 2. Assay conditions:
Wavelength 340nm Wavelength 340 nm
Temperature 37 oC Temperature 37 oC
Cuvette 1cm light path Cuvette 1cm light path
Intended Use 3. Adjust the instrument to zero with distilled water . Intended Use 3. Adjust the instrument to zero with distilled water .
In vitro diagnostic reagents for the quantitative determination of 4. Pipette into a cuvette : In vitro diagnostic reagents for the quantitative determination of 4. Pipette into a cuvette
Kappa Light Chain (KAP) in human serum by turbidimetric Kappa Light Chain (KAP) in human urine by turbidimetric
immunoassay. immunoassay.
Standard Sample Standard Sample
Background Reagent (R1) 350 ml 350 ml Background Reagent (R1) 250 ml 250 ml
The determination of Kappa/Lambda in human serum is important Standard 2 ml ----- The presence of monoclonal free light chains,i.e.Bence jones Standard 4 ml -----
for the diagnosis and subtyping of momoclonal gammopathies. proteins(BJ) in urine is of great importance as an aid in the diagnosis
Where as polyclonal immunoglobulins(normal or enhanced Sample ----- 2 ml of B cell malignancies such as multiple myeloma and non-Hodgkin Sample ----- 4 ml
concentration) exhibit both Kappa and Lambda types of light chains Mix and incubate for 2 minutes at 37 C, o lymphoma,and in monitoring their therapy. Mix and incubate for 2 minutes at 37 C, o
in a roughly constant ratio of 2:1 ,monoclonal immunoglobulins read absorbance (A1). Where as complete immmunoglobulin molecules can not pass the read absorbance (A1).
exhibit only one type of light chain. intact glomerular filtiration barrier,free immunoglobulin light chains
Increased production of monoclonal immunoglobulins or free Reagent (R2) 75 ml 75 ml are filtered through the glomeruli and reabsorbed in the tubuli. Reagent (R2) 30 ml 30 ml
monoclonal light chains leads to Kappa/Lambda quotient outside After addition of R2, incubate and after 5 minutes at 37 C and o Elevated concentrations of free Kappa light chain in serum,such as After addition of R2, incubate and after 5 minutes at 37 oC and
the reference range indicating the existence of a monoclonal record 2nd reading (A2). are realeased by monoclonal plasma cells,may exceed the tubular record 2nd reading (A2)
gammopathy. reabsorption capacity and lead to the excretion of free Kappa light
Calculation chains in the urine. Calculation
Test Principle Generate a reference curve by successive 1 : 2 dilutions of Generate a reference curve by successive 1 : 2 dilutions of
Kappa Light Chain (KAP) test is based upon the antigen-antibody Standard (Protein Standard high) in saline (At Least 5 points are Test Principle Standard (SPecific Protein Standard) in saline (At Least 5 points are
reaction. recommended). Use Saline as zero point. Determine D absorbance Kappa Light Chain (KAP) test is based upon the antigen-antibody recommended). Use Saline as zero point. Determine D absorbance
of the sample and each standard as following: reaction by the end point method. of the sample and each standard as following:
Reagents D absorbance of sample = (A2 - A1) sample D absorbance of sample = (A2 - A1) sample
R1 Buffer Reagent D absorbance of each standard = (A2 - A1) for each standard Reagents D absorbance of each standard = (A2 - A1) for each standard
Phosphate buffered saline(pH 7.43). Plot the calibration curve and obtain the result. R1 Buffer Reagent Plot the calibration curve and obtain the result.
Enhancer. Phosphate buffered saline(pH 7.43).
Sodium azide (0.95 g/L). Sensitivity Enhancer. Linearity
40 mg/dL. Sodium azide (0.95 g/L). The reaction is linear up to concentration of 340 mg/L.; specimens
R2 Antiserum showing higher concentration should be diluted 1+2 with physiological
Phosphate beffered saline(pH 7.43). Linearity R2 Antiserum saline and repeat the assay (result×3).
Polyclonal goat anti-human Kappa Light Chain (variable). The reaction is linear up to concentration of 800 mg/dL.; specimens Phosphate beffered saline(pH 7.43).
Sodium azide (0.95 g/L). showing higher concentration should be diluted 1+2 with physiological Polyclonal goat anti-human Kappa Light Chain (variable). Expected Values
saline and repeat the assay (result×3). Sodium azide (0.95 g/L). < 10 mg/L .
Materials required but not provided with the kit: Each laboratory should establish an expected range for the
1. Sodium chloride (9 g/l) Expected Values Materials required but not provided with the kit: geographical area in which it is located
2. Standard (Protein standard high (REF 604 001) is required for 200 - 440 mg/dL (IFCC). 1. Sodium chloride (9 g/l)
constructing calibration curve). 2. Standard (Specific Protein standard (REF 387 001) is required References
3. controls. ( Immunology control low (REF 606 001) and Each laboratory should establish an expected range for the for constructing calibration curve). 1. Tillyer,C.R,Int.J.Clin.Lab.Res.22,152 (1982).
immunology control high (REF 608 001) is required for quality geographical area in which it is located. 3. controls. ( Specific Protein control (REF 390 001) control is 2. Boege, F. et al, J.Clin.Chem.Clin.Biochem. 28, 37 (1990).
control. required for quality control. 3. Tillyer,C.R,Int.J.Clin.Lab.Res.23,25 (1993).
References
Precautions and Warnings 1. Boege, F. et al, Lab. Med. 13, 369-374 (1989) Precautions and Warnings
1. For in vitro diagnostic use only. 2. Lievens, M. M, J. Clin. Chem. Clin.Biochem.27, 519-523 (1989). 1. For in vitro diagnostic use only.
2. Sodium azide has been reported to form lead or copper azides 3. Dati, F. et al, Lab. Med. 13, 87-90 (1989). 2. Sodium azide has been reported to form lead or copper azides ORDERING INFORMATION
in laboratory plumbing which may explode on percussion.Flush in laboratory plumbing which may explode on percussion.Flush
drains with water thoroughly after diposing of fluids containing drains with water thoroughly after diposing of fluids containing CATALOG NO QUANTITY
sodium azide. sodium azide.
ORDERING INFORMATION 378 001 50 test
Reagents Preparation, Storage and Stability CATALOG NO QUANTITY Reagents Preparation, Storage and Stability
Reagents are liquid, ready to use. Reagents are liquid, ready to use.
Reagents are stable until the expiration date when kept at (2 - 8 ºC). 375 001 50 test Reagents are stable until the expiration date when kept at (2 - 8 ºC).
Stability in the instrument is at least 4 weeks if contamination is Stability in the instrument is at least 4 weeks if contamination is
avoided. DO NOT FREEZE. avoided. DO NOT FREEZE.
Specimen Collection and Preparation Specimen Collection and Preparation

Use fresh serum, If the test can not be carried out on the same day Use fresh centrifugated urine, If the test can not be carried out on
the serum may be stored at 2 - 8 ºC for 48 hours. the same day ,the urine may be stored at 2 - 8 ºC for 48 hours.
If stored for longer period the sample should be frozen. If stored for longer period the sample should be frozen.
210 211
Lambda light chain serum kit SYMBOLS IN PRODUCT LABELLING Lambda light chain urine kit SYMBOLS IN PRODUCT LABELLING
(LAM) EC REP Authorised Representative

(LAM) EC REP Authorised Representative
Quantitative determination of Lambda light chain (LAM) REF Catalogue Number for use
in human serum by turbidimetric immunoassay. Consult instructions for use Manufactured by human urine by turbidimetric immunoassay. Consult instructions for use Manufactured by
REF: 381 001 50 test REF: 384 001 50 test

R1 Buffer Reagent 1 x 18 ml Procedure R2 Antiserum 1 X1.7 ml
R2 Antiserum 1 X 4 ml 1. Bring the reagents to 37 oC Procedure
2. Assay conditions: 1. Bring the reagents to 37 oC
Wavelength 340 nm 2. Assay conditions:
Temperature 37 oC Wavelength 340 nm
Cuvette 1cm light path Temperature 37 oC
Intended Use 3. Adjust the instrument to zero with distilled water . Intended Use Cuvette 1cm light path
In vitro diagnostic reagents for the quantitative determination of 4. Pipette into a cuvette : In vitro diagnostic reagents for the quantitative determination of 3. Adjust the instrument to zero with distilled water .
Lambda Light Chain (LAM) in human serum by turbidimetric Lambda Light Chain (LAM) in human urine by turbidimetric 4. Pipette into a cuvette
immunoassay. immunoassay.
Standard Sample
Standard Sample
Background Reagent (R1) 350 ml 350 ml Background
The determination of Kappa/Lambda in human serum is important The presence of monoclonal free light chains,i.e.Bence jones Reagent (R1) 250 ml 250 ml
Standard 2 ml -----
for the diagnosis and subtyping of momoclonal gammopathies. proteins(BJ) in urine is of great importance as an aid in the diagnosis Standard 8 ml -----
Where as polyclonal immunoglobulins (normal or enhanced Sample ----- 2 ml of B cell malignancies such as multiple myeloma and non-Hodgkin
concentration) exhibit both Kappa and Lambda types of light chains lymphoma,and in monitoring their therapy. Sample ----- 8 ml
Mix and incubate for 2 minutes at 37 oC,
in a roughly constant ratio of 2:1 ,monoclonal immunoglobulins read absorbance (A1). Where as complete immmunoglobulin molecules can not pass the Mix and incubate for 2 minutes at 37 C, o
exhibit only one type of light chain. intact glomerular filtiration barrier,free immunoglobulin light chains read absorbance (A1).
Increased production of monoclonal immunoglobulins or free Reagent (R2) 75 ml 75 ml are filtered through the glomeruli and reabsorbed in the tubuli.
monoclonal light chains leads to Kappa/Lambda quotient outside Elevated concentrations of free Kappa light chain in serum,such as Reagent (R2) 30 ml 30 ml
After addition of R2, incubate and after 5 minutes at 37 oC and
the reference range indicating the existence of a monoclonal record 2nd reading (A2). are realeased by monoclonal plasma cells,may exceed the tubular After addition of R2, incubate and after 5 minutes at 37 oC and
gammopathy. reabsorption capacity and lead to the excretion of free Kappa light record 2nd reading (A2)
Calculation chains in the urine.
Test Principle Generate a reference curve by successive 1 : 2 dilutions of Calculation
Lambda Light Chain (LAM) test is based upon the antigen-antibody Standard (Protein standard high) in saline (At Least 4 points are Test Principle Generate a reference curve by successive 1 : 2 dilutions of Standard
reaction. recommended). Use Saline as zero point. Determine D absorbance Lambda Light Chain (LAM) test is based upon the antigen-antibody in saline (At Least 4 points are recommended). Use Saline as zero
of the sample and each Standard as following: reaction by the end point method. point. Determine D absorbance of the sample and each Standard
Reagents D absorbance of sample = (A2 - A1) sample as following:
R1 Buffer Reagent D absorbance of each Standard = (A2 - A1) for each standard Reagents D absorbance of sample = (A2 - A1) sample
Phosphate buffered saline ( pH 7.43). Plot the calibration curve and obtain the result. D absorbance of each Standard = (A2 - A1) for each Standard
Enhancer. R1 Buffer Reagent Plot the calibration curve and obtain the result.
Sodium azide (0.95 g/L). Sensitivity Phosphate buffered saline ( pH 7.43).
20 mg / dL. Enhancer. Linearity
R2 Antiserum Sodium azide (0.95 g/L). The reaction is linear up to concentration of 200 mg/L.; specimens
Phosphate beffered saline (pH 7.43). Linearity showing higher concentration should be diluted 1+2 with physiological
Polyclonal goat anti-human Lambda Light Chain (variable). The reaction is linear up to concentration of 450 mg/dL.; specimens R2 Antiserum saline and repeat the assay (result×3).
Sodium azide (0.95 g/L). showing higher concentration should be diluted 1+2 with physiological Phosphate beffered saline (pH 7.43).
saline and repeat the assay (result×3). Polyclonal goat anti-human Lambda Light Chain (variable). Expected Values
Materials required but not provided with the kit: Sodium azide (0.95 g/L). < 10 mg/L .
1. Sodium chloride (9 g/l) Expected Values
2. Standard (Protein standard high (REF 604 001) is required for 110 - 240 mg/dL .(IFCC) Materials required but not provided with the kit: Each laboratory should establish an expected range for the
constructing calibration curve). 1. Sodium chloride (9 g/l) geographical area in which it is located.
3. controls. ( Immunology control low (REF 606 001) and Each laboratory should establish an expected range for the 2. Standard (Specific Protein standard (REF 387 001) is required
immunology control high (REF 608 001) is required for quality geographical area in which it is located. for constructing calibration curve). References
control. 3. controls. ( Specific Protein control (REF 390 001) control is 1. Tillyer,C.R,Int.J.Clin.Lab.Res.22,152 (1982).
References required for quality control. 2. Boege, F. et al, J.Clin.Chem.Clin.Biochem. 28, 37 (1990).
Precautions and Warnings 1. Boege, F. et al, Lab. Med. 13, 369-374 (1989) 3. Tillyer,C.R,Int.J.Clin.Lab.Res.23,25 (1993).
1. For in vitro diagnostic use only. 2. Lievens, M. M, J. Clin. Chem. Clin.Biochem.27, 519-523 (1989). Precautions and Warnings
2. Sodium azide has been reported to form lead or copper azides 3. Dati, F. et al, Lab. Med. 13, 87-90 (1989). 1. For in vitro diagnostic use only.
in laboratory plumbing which may explode on percussion.Flush 2. Sodium azide has been reported to form lead or copper azides
drains with water thoroughly after diposing of fluids containing in laboratory plumbing which may explode on percussion.Flush
sodium azide. drains with water thoroughly after diposing of fluids containing ORDERING INFORMATION
ORDERING INFORMATION sodium azide. CATALOG NO QUANTITY
Reagents Preparation, Storage and Stability CATALOG NO QUANTITY
Reagents are liquid, ready to use. Reagent Preparation, Storage and Stability 384 001 50 test
Reagents are stable until the expiration date when kept at (2 - 8 ºC). 381 001 50 test Reagents are liquid, ready to use.
Stability in the instrument is at least 4 weeks if contamination is Reagents are stable until the expiration date when kept at (2 - 8 ºC).
avoided. DO NOT FREEZE. Stability in the instrument is at least 4 weeks if contamination is
avoided. DO NOT FREEZE.
Use fresh serum, If the test can not be carried out on the same day Specimen Collection and Preparation
the serum may be stored at 2 - 8 ºC for 48 hours. Use fresh centrifuged urine , If the test can not be carried out on the
If stored for longer period the sample should be frozen. same day, the urine may be stored at 2 - 8 ºC for 48 hours.
If stored for longer period the sample should be frozen.
212 213
LIPOPROTEIN (a) [Lp(a)] SYMBOLS IN PRODUCT LABELLING
Calibration curve
References
Calibrator 1 100 ml of Spectrum LP(a) Calibrator* 1. Berg K. A new serum type system in man: The Lp-system. Acta
Calibrator 2 100 ml of Calibrator 1 + 100 ml of Saline Solution Pathol. Microbiol. Scand. 1963;59:369-82.
Calibrator 3 100 ml of Calibrator 2 + 100 ml of Saline Solution 2. Gaubatz JW, Heideman C, Gotto A MJr., Morriset JD, Dahlen
REF: 550 001 50 test REF
Consult instructions for use Manufactured by Calibrator 4 100 ml of Calibrator 3 + 100 ml of Saline Solution G.H. Human plasma lipoprotein (a): Structure and properties.
R1 Buffer 1X 22 ml
Calibrator 5 100 ml of Saline Solution J.Biol. Chem.1983;258: 4582-89.
R2 Latex 1 X 3.2 ml
(*) See values on the label or on the insert. Multiply by the 3. Scanu A M, Fless G M. Lipoprotein (a). J. Clin. Invest.1990;
All raw materials of human origin used in the manufacture of this appropriate factor. 85:1709-15.
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 4. Sandkamp M, FunkeH, Schulte H, Kohler E, Assmann G.
Intended Use and HCV with commercially available test methods. However, For quality control use Spectrum Control or other suitable control Lipoprotein (a) is an independent risk factor for myocardial
In vitro diagnostic reagents for the quantitative determination this product should be handled as though capable of transmitting material. The control intervals and limits must be adapted to the infarction at a young age. Clin Chem.1990;36: 20-3.
of Lipoprotein (a) [Lp(a)] in human serum by means of particle- infectious diseases individual laboratory requirements. Values obtained should fall 5. Rosengren A, Wilhelmsen L, Eriksson E, Risberg B, Wedel
enhanced turbidimetric immunoassay. within established limits. Each laboratory should establish corrective H.Lipoprotein (a) and coronary heart disease: A prospective
Precautions and Warnings measures to be taken if values fall outside the limits. Control must be case-control study in a general population sample of middle
age men. Br. Med. J.1990; 301: 1248-51.
Background For in vitro diagnostic use only. Do not pipette by mouth. Reagents assayed and evaluated as for patient samples.
Lipoprotein (a) [Lp(a)] was initially though to be a genetic variant containing sodium azide must be handled with precaution. Sodium 6. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
Edition, AACC Press, 2000.
of low density lipoprotein (LDL). Lp(a) is a low density lipoprotein- azide can form explosive azides with lead and copper plumbing. Procedure
like particle containing apolipoprotein B-100 disulphide-linked to Since absence of infectious agents cannot be proven, all specimens Wavelength 600 nm 7. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
one large glycoprotein called apolipoprotein (a). Apolipoprotein (a) and reagents obtained from human blood should always be handled Temperature 37 ºC 207 – 224
has been shown to have a considerable degree of homology with with precaution using established good laboratory practices. Cuvette 1cm light path
human plasminogen. The characteristic feature of lipoprotein (a) is Disposal of all waste material should be in accordance with local Measurement against distilled water blank.
that it is distinct from all other serum proteins and apolipoproteins. guidelines. Bring the reagents at 37ºC and pipette: ORDERING INFORMATION
This protein is believed to be inherited as an autosomal dominant As with other diagnostic tests, results should be interpreted Blank Calibrator Sample CATALOG NO QUANTITY
trait and appears to be insensitive to either diet, lifestyle or most considering all other test results and the clinical situation of the Work. Reagent 480 ml 480 ml 480 ml
hypolipidaemic drugs. patient. 550 001 50 test
Distilled Water 4 µl ------ ------
Since its discovery by Berg in 1963, there has been a considerable
Calibrator ------ 4 µl ------
rise in interest, not only in specialized research centres but also Material Required
in clinical routine laboratories, in the accurate measurement of Spectrophotometric analyzer. Sample ------ ------ 4 µl
lipoprotein (a) in blood. This interest was stimulated by reports Saline solution. Mix and measure absorbance immediately (A1) incubate 4
indicating that levels above 0,2 . 0,3 g/l, present in approximately Controls. min (37ºC), after incubation read absorbance (A2).
25 % of the population, are associated with an increased risk of
coronary heart disease. Many investigators have confirmed that a Storage and Stability Calculation
high lipoprotein(a) concentration represents an indicator of risk for The Lp(a) reagents should be stored tightly capped at (2 - 8 ºC) when Determine D absorbance of the sample and each calibrator as
cardiovascular disease, especially when the serum LDL-cholesterol not in use. Do not freeze. Reagents in the original vials are stable following:
or apo B are elevated. Therefore a convenient and reliable method to the expiration date on the vial label when capped and stored at D absorbance of sample = (A2 - A1) sample
for the quantitation of Lp(a) in serum or plasma is important for (2 - 8 ºC). Immediately following the completion of an assay run, the D absorbance of each calibrator = (A2 - A1) for each calibrator
identification of individuals at risk for developing atherosclerosis. reagent vials should be capped until next use in order to maximize Plot the calibration curve and obtain the result.
curve stability. Once opened the reagent can be used within 1 month If is an one point calibration
Test Principle if stored tightly closed at (2 - 8 ºC) after use.
This Lp(a) test is based upon the reactions between Lp(a) in The Lp(a) buffer reagent should be clear and colourless. Any turbidity x Calibrator concentration
the sample and latex-covalently bound rabbit antihuman Lp(a) may be sign of deterioration and reagent should be discarted. (A2-A1) calibrator - (A2-A1) blank
antibodies. Lp(a) values are determined photometrically. The Lp(a) latex reagent should have a white, turbid appearance free
of granular particulate. Sensetivity
Reagents Visible agglutination or precipitation may be a sign of deterioration, < 15 mg/L
Buffer R1 and the reagent should be discarted.
Glycine buffer, pH: 8.0, containing protein stabilizers and 0.09 % Linearity
sodium azide as preservative. Specimen Collection and Preparation Up to 800 mg/L.
Latex reagent R2
good laboratory practices. Expected Values
a suspension of latex microparticles covalently bound antibodies
against human Lp(a) in a glycine buffer (0.1 M, pH: 8,2), contain- Lp(a) remain stable for 14 days at (2 - 8 ºC). if the test should be Values < 300 mg/L are within the normal range. This data must be
ing NaCL (0.15M) and bovine serum albumin (0.5%). Preservative: performed later, it is recommended to freeze the serum. Lipemic interpreted as a guide.
Sodium azide 0.075%.. specimens, or turbid specimens, must be clarified before the assay Each laboratory should establish an expected range for the
by high-speed centrifugation (10 min at approx. 15.000 rpm). geographical area in which it is located.
Calibrator
Human - based reference fluid. Preservative: sodium azide, 0.075%.
reconstitute the lyophilyzed calibrator vial with the amount of distilled Reagent Preparation
water stated on the vial. Working Reagent is prepared with 1 part of Latex Reagent and 7
Stability 2 weeks after reconstitution when stored at 2-8 ºC. parts of Buffer Reagent. Prepare a fresh WR based on its workload.
Shake gently the reagents before pipetting.
e.g: 60 ml of latex reagent& 420 ml of buffer reagent.
214 215
Microalbumin Standard SYMBOLS IN PRODUCT LABELLING
REF 352 001 1x1 ml
Specifications
Lot number : MAL 3014
Expiration Date : July 2016
Source : Human plasma
o
Storage: 2-8 C Do Not Freeze
Stabilitty after opening the vial : 6 weeks
Target MAL mg/L

Microalbumin Standard
432
216 217
MICROALBUMIN (MAU) SYMBOLS IN PRODUCT LABELLING
Calculation
Generate a reference curve by successive 1 : 2 dilutions of standard
Immuno Turbidimetry EC REP Authorised Representative
LOT Batch Code/Lot number CAUTION. Consult instructions following:
REF Catalogue Number for use D absorbance of sample = (A2 - A1) sample
D absorbance of each calibrator = (A2 - A1) for each standard
REF: 600 001 100 test Plot the calibration curve and obtain the result
R1 Buffer 2 x 20 ml
R2 Antiserum 1 X 6 ml Sensitivity
0.7 mg/L
Precautions and Warnings Linearity

Intended Use For in vitro diagnostic use only. Do not pipette by mouth. Reagents 400 mg/L
In vitro diagnostic reagents for the quantitative determination of Mi- containing sodium azide must be handled with precaution. Sodium
croalbumin (MAU) in urine by means of particle-enhanced turbidi- azide can form explosive azides with lead and copper plumbing. Quality Control
metric immunoassay in clinical chemistry analyzers. Since absence of infectious agents cannot be proven, all specimens Control sera are recommended to monitor the perfomance of manual
and reagents obtained from human blood should always be handled and automated assay procedures .
Background with precaution using established good laboratory practices. Each laboratory should establish its own Quality Control Scheme
Increased albumin excretion detectable only by sensitive and corrective actions if controls do not meet the acceptable
immunoassay (microalbuminuria) has been used for some years as Disposal of all waste material should be in accordance with local tolerances.
a predictor of incipient nephropathy and cardiovascular disease in guidelines.
diabetic patients. Microalburninuria has also been associated with Expected Values
hypertension and increased risk of cardiovascular disease in non- As with other diagnostic tests, results should be interpreted consid-
0-25 mg/L
diabetic patients. Microalbuininuria occurs in response to acute ering all other test results and the clinical situation of the patient.
inflammatory conditions such as ischaemia, trauma,and termal Each laboratory should establish an expected range for the
injury, surgery, pancreatitis, and inflammatory bowel disease. Reagent Preparation , Storage and Stability geographical area in which it is located.
In many of these conditions albumin excretion increases within All reagents are supplied ready to use.
minutes or hours of the initiating stimulus and only last for 24 to 72 h. Reagents in the original vial are stable to the expiration date on the
References
The degree of microalburninuria is proportional to the severity of the vial label when capped and stored at (2 - 8 ºC).
1. Medcalf E A et al. Clin Chem 1990; 36/3: 446-449.
inflammatory insult, is predictive of outcome, and is not associated
2. Mount,J.N.J. Chin pathology,22,12(1986)
with any other features of renal impairment. Conventional dip-stick Standard: 3. Panuyiotou B N. Journal International Medical Research 1994;
and acid precipitation tests for detecting protein in urine lack the The standard is stable to the expiration date on the vial label when 22: 181-201.
sensitivity required to delineate this condition. Dip-stick may yield capped and stored at (2 - 8 ºC). 4. Shmidtz,A,et al, Diabetic Medicine,5,126(1988).
negative or trace results even when the albumin excretion rate is Once opened the standard is stable for 6 weeks if stored tightly
10 or 20 times normal; and the rate must increase to 200 or 300 closed at 2 - 8 ºC after use.
micrograms per minute (mg/min) before nephropathy becomes
clinically apparent as persistent proteinuria. Interest in measuring For further storage at - 30 ºC divide standard into aliquots. Once
subclinical elevations in the albumin excretion rate has focused thawed never freeze again. ORDERING INFORMATION
on individuals with an already established diagnosis of diabetes or
essential hypertension. Providing proper care is taken to minimise CATALOG NO QUANTITY
the influence of exercise and poor metabolic control of the albumin Collect urine during 24 hours or as a random midstream
excretion rate, the urinary albumin level has proved to be an 600 001 100 test
sample.If the test can not be carried out on the same day,the urine
excellent predictor of the progression to overt nephropaty in both may be stored at (2 - 8 ºC )for 48 hours.If stored for a longer
insulin-dependent and non-insulin dependent diabetes. period,the sample should be frozen.
The use of centrifuged urine is recommended.
Test Principle
This MAU test is based upon the MAU antigen-antibody reaction. Procedure
1. Bring the reagents and the photometer to room temperature
2. Assay conditions:
Temperature room temperature
R1 Buffer Cuvette 1cm light path
saline(9 g/L). 3. Adjust the instrument to zero with distilled water .
Accelerator. 4. Pipette into a cuvette
Sodium azide (0.95 g/L)
standard Sample
R2 Antiserum Reagent (R1) 400 mL 400 mL
phosphate buffered saline.
Polyclonal goat anti-human Albumin(variable).
Sample ------ 25 ml
Mix and record 1st reading (A1).
Materials required but not provided with the kit Reagent (R2) 60 ml 60 ml
1- Standard
Microalbumin concentration is stated on the vial label. After addition of R2, incubate at room temperature and after 5
minutes record 2nd reading (A2)
2-Controls
218 219
MICROALBUMIN (MAU) SYMBOLS IN PRODUCT LABELLING
Calibration
Use Microalbumin Calibrator. Expected Values
Turbi Latex EC REP Authorised Representative Temperature Limitation The sensitivity of the assay and the target value of the calibrator Up to 30 mg/24 hrs urine specimen.
IVD For in-vitro diagnostic use Use by/Expiration Date have been standardized against the International Reference Material 20 mg/L in a first morning urine specimen.
CRM 470/RPPHS.The on board calibration is stable for 3 weeks. Each laboratory should establish its own reference range.
REF: 563 001 100 test •Recalibrate when control results are out of specified tolerances,
R1 Buffer 2 x 20 ml when using different lot of reagent and when the instrument is References
R2 Latex 1 X 10 ml adjusted. 1. Medcalf E A et al. Clin Chem 1990; 36/3: 446-449.
2. Panuyiotou B N. Journal International Medical Research 1994;
Quality Control
22: 181-201.
Intended Use Calibrator Control sera are recommended to monitor the perfomance of manual
3. Feldt-Rasmussen B et al. J Diab Comp 1994; 8: 137-145.
In vitro diagnostic reagents for the quantitative determination Liquid Calibrator. Microalbumin concentration is stated on the vial and automated assay procedures .
of Microalbumin (MAU) in urine by means of particle-enhanced label. Each laboratory should establish its own Quality Control Scheme 4. Gilbert R E et al. Diabetic Medicine 1994; 11: 636-645.
turbidimetric immunoassay in clinical chemistry analyzers. and corrective actions if controls do not meet the acceptable 5. Bar J et al. Diabetic Medicine 1995; 12: 649-656.
Precautions and Warnings tolerances.
6. Young DS. Effects of drugs on clinical laboratory test, 4th ed.
Background For in vitro diagnostic use only. Do not pipette by mouth. Reagents AACC Press, 1995.
Increased albumin excretion detectable only by sensitive containing sodium azide must be handled with precaution. Sodium Procedure
immunoassay (microalbuminuria) has been used for some years as azide can form explosive azides with lead and copper plumbing. 1. Bring the reagents and the photometer to 37ºC
a predictor of incipient nephropathy and cardiovascular disease in Since absence of infectious agents cannot be proven, all specimens 2. Assay conditions:
diabetic patients. Microalburninuria has also been associated with and reagents obtained from human blood should always be handled Wavelength 540 nm (530 -550 nm)
hypertension and increased risk of cardiovascular disease in non- with precaution using established good laboratory practices. Temperature 37ºC CATALOG NO QUANTITY
diabetic patients. Microalbuininuria occurs in response to acute Cuvette 1cm light path
563 001 100 test
inflammatory conditions such as ischaemia, trauma,and termal Disposal of all waste material should be in accordance with local 3. Adjust the instrument to zero with distilled water .
injury, surgery, pancreatitis, and inflammatory bowel disease. In guidelines. 4. Pipette into a cuvette :
many of Working Reagent 500 ml
these conditions albumin excretion increases within minutes or hours As with other diagnostic tests, results should be interpreted Calibrator or Sample 5 ml
of the initiating stimulus and only last for 24 to 72 h. The degree of considering all other test results and the clinical situation of the 5.Mix and read absorbance immediately (A1) and after 2 minutes
microalburninuria is proportional to the severity of the inflammatory patient. (A2) of the sample addition.
insult, is predictive of outcome, and is not associated with any other
features of renal impairment. Conventional dip-stick and acid Material Required Calculation
precipitation tests for detecting protein in urine lack the sensitivity Spectrophotometric analyzer. Plot the calibration curve and the sample concentration is obtained
required to delineate this condition. Dip-stick may yield negative or Thermostatic bath at 37ºC. by interpolation the sample absorbance (A2-A1) in the calibration
trace results even when the albumin excretion rate is 10 or 20 times curve.
normal; and the rate must increase to 200 or 300 micrograms per Storage and Stability If is an one point calibration:
minute (mg/min) before nephropathy becomes clinically apparent as Reagents in the original vial is stable to the expiration date on the vial
(A2-A1) sample
persistent proteinuria. Interest in measuring subclinical elevations label when capped and stored at (2 - 8 ºC). Immediately following the mg/L (MAU) = x Calibrator concentration
in the albumin excretion rate has focused on individuals with an completion of an assay run, the reagent vial should be capped until (A2-A1) calibrator
already established diagnosis of diabetes or essential hypertension. next use and contaminations are prevented during their use. Do not
Providing proper care is taken to minimise the influence of exercise use reagents over the expiration date. For conversion to mg/min:
and poor metabolic control of the albumin excretion rate, the The MAU latex reagent should have a white, turbid appearance free
urinary albumin level has proved to be an excellent predictor of the of granular particulate. Visible agglutination or precipitation may be a mg Microalbumin in 24hrs Urine
progression to overt nephropaty in both insulin-dependent and non- sign of deterioration, and the reagent should be discarted. The MAU mg/min (Microalbumin) =
insulin dependent diabetes. diluent should be clear and colourless. Any turbidity may be sign of 1440
deterioration and reagent should be discarted. WR is stable for up to where 1440 = 24 x 60 (minutes /day)
Test Principle one day at 2-8ºC. It is recommended that each Laboratory prepares
This MAU test is based upon the reactions between albumin and a fresh Working Reagent based on its workload. example:
latex-covalently bound antibodies against human albumin. MAU - If 24hrs urine is 1.8 L (1800 ml)
values are determined photometrically. Specimen Collection and Preparation - Microalbumin concentration is 15 mg/L
24 hours or random/ first morning urine specimen. It is recommended
Reagents to adjust the pH at 7.0 with NaOH/HCL 1 mol/L.Stable 7 days at Microalbumin Concentration in mg/day = 15 x 1000 x 1.8
2-8ºC when sodium azide 1 g/L is added to prevent contamination. = 27000 mg
R1 Buffer Urine should be centrifuged before testing. *where 1000 is to convert from mg/L to mg/L
Glycine buffer 100 mmol/L, pH 10.0. Sodium azide 0.90 g/L. 1.8 is the volume of 24hrs Urine.
Merthiolate 0.05 g/L. Reagent Preparation and Stability
Working Reagent (WR) is prepared with 1 part of Latex Reagent and 27000
R2 Latex reagent 4 parts of Buffer. Prepare a fresh WR based on its workload. Shake mg/min (Microalbumin) = = 18.7
Particles coated goat IgG with anti -human albumin, pH 8.2. Sodium gently the reagents before pipetting. 1440
azide 0.90 g/L. Merthiolate 0.05 g/L. Linearity
Stability: WR is stable for up to one day at 2-8ºC. Up to 150 mg/L
220 221
Microalbumin Control set SYMBOLS IN PRODUCT LABELLING Protein standard High SYMBOLS IN PRODUCT LABELLING
IVD For in-vitro diagnostic use Use by/Expiration Date IVD For in-vitro diagnostic use Use by/Expiration Date
REF: 604 001 1x1 ml
REF: 353 001 2 x 1 ml REF: 604 002 2x1 ml
Intended Use
Intended Use Accuracy control for the determination of protiens in serum by turbidimetry and nephelometry.
Accuracy control for the determination of Microalbumin in serum by turbidimetry and nephelometry
composition
Composition A dilution of humen plasma with phosphsate buffered saline.
A dilution of humen plasma containing high levels of Microalbumin with phosphsate buffered saline. Contains stabilizers and 0.095 g% sodium azide .
Preservative: 0.095 g% sodium azide .
Storage and Stability The expiry date of the product at +2 to +8 ºC is listed on the label.
The expiry date of the product at +2 to +8 ºC is listed on the label. After first opening the standard can be used for 6 weeks if stored tightly closed
After first opening the control can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use.
at +2 to+8 ºC after use.
Do not freeze.
Do not freeze.
Precautions and Warnings 1. For “In Vitro Diagnosticc Use”
1. For “In Vitro Diagnostic Use” 2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin
2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin (HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were
(HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in
used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in accordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven .
accordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven . 3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes !
Lot No.: MPH021
Assigned Value
1- Lot number : MALL 1014 Protein Protein Values (mg/dl)

Albumin 11812
Microalbumin : 213 ( 192 - 234 ) mg/L.
a1-Antitrypsin 410
2- Lot number : MALH 1014 a2-Macroglobulin 665
Microalbumin : 426 (400 - 452 ) mg/L a1-Acidglycoportein 225
Ceruloplasmin 90
Haptoglobin 355
Transferrin 797
IgA 574
IgG 2632
IgM 359
Complement C3 432
Complement C4 72
Antithrombin III 74
C1 esterase inhibitor 77
Kappa Light chain 654
Lambada light chain 381
Prealbumin 74
Values based on a reference preparation (ERM-DA470K/IFCC)from the International Federation of clinical chemistry(IFCC).
values based on siemens standard material.
222 223
Procalcitonin Assay SYMBOLS IN PRODUCT LABELLING
Calibration
The Spectrum PCT Assay should be calibrated using Spectrum
Limitation
Samples with PCT level exceeding the linearity limit of 60 ng/mL
EC REP Authorised Representative Use by/Expiration Date PCT Calibrator PCT concentration in sample is determined from a should be diluted with negative sample. Rerun the tests and multiply
IVD For in-vitro diagnostic use ! CAUTION. Consult instructions calibration curve obtained from the PCT Calibrators . The PCT the results by the dilution factors. As with any latex turbidimetric
LOT Batch Code/Lot number for use Calibrators are lyophilized powder and require reconstitution with immunoassays, Diazyme PCT Assay runs should be followed with
REF: 567 001 50 test DI water before use. appropriate and thorough wash steps. Please consult specific
R1 buffer 1 x 14 ml calibration frequency Biweekly calibration is recommended instrument manuals for further information.
R2 Latex 1 x 4.7 ml Temperature Limitation
Quality Control References
Good laboratory practice recommends the use of control materials. 1. Müller B, et al., Ubiquitous expression of the calcitonin-i genein
Users should follow the appropriate federal, state and local guideline multipletissues in response to sepsis. J Clin Endocrinol Metab
Intended Use Reagent Preparation, Storage and Stability concerning the running of external quality control.To ensure adequate 2001; 86(1):396-404.
Spectrum Procalcitonin Assay is for the quantitative determination The reagents are ready-to-use and stable when stored at 2 – 8 oC. quality control, normal and abnormal controls with known values 2. Christ-Crain M, et al., Procalcitonin in bacterial infections
of Procalcitonin Assay(PCT) in serum samples, EDTA or lithium until the expiry date labeled on the bottles should be run as unknown samples.PCT Control are lyophilized hype, hopehope or more or less? Swiss Med Wkly 2005;
heparin plasma samples by latex enhanced immunoturbidimetric powder and require reconstitution with cold DI water before use. 135:45160.
method. DO NOT FREEZE 3. Schuetz P, et al., Long-term stability of procalcitonin in frozen
Linearity samples and comparison of Kryptor and VIDAS automated
Background Specimen Collection and Preservation The linearity of the assay is from 0.17-60 ng/mL. Results that exceed immunoassays. Clin Biochem.2010;43(3):3414
Procalcitonin (PCT) is a 116 amino acid protein, the prohormone of The PCT Assay is formulated for use with serum or lithium heparin 60 ng/mL. should be diluted with saline and retested. 4. Stolz D, Christ-Crain M, Bingisser R, Leuppi J, Miedinger
calcitonin. and EDTA plasma. For monitoring purpose, the same sample D,MullerC, Huber P, Muller B, Tamm M. Antibiotic treatment
Whereas hormonally active calcitonin is produced exclusively in the matrix should always be used. PCT values measured in arterial of exacerbations of COPD: a randomized, controlled trial
C-cells of the thyroid gland after specific intracellular proteolytic blood are about 4% higher than from venous blood. PCT increases comparing procalcitonin-guidance with standard therapy.
The substances normally present in the plasma were tested.
procession of the prohormone PCT, PCT is ubiquitously and about 3 hours after bacterial infection, Chest. 2007;131:9-19.
Less than 10% deviation was produced when tested up to the
uniformly expressed in multiple tissues throughout the body in reaching maximum values after 6-12 hours with a half life of 25 to
concentrations shown below
response to sepsis. In healthy conditions, the PCT levels in the 30 hours.
Ascorbic Acid 10 mM
circulation are very low (< 0.05 ng/ml). Elevated circulating levels of PCT is relative stable in both plasma and serum samples, and
Bilirubin, free 40 mg/dL
PCT are important indicators in response to microbial infections and no special requirements for pre-analytical sample handling.
Bilirubin, conjugated 40 mg/dL ORDERING INFORMATION
a powerful tool in the early detection of sepsis
Hemoglobin 400 mg/dL
Samples stored at 4°C retain CATALOG NO QUANTITY
Triglyceride 1000 mg/dL
Method >90 percent for several days, whereas samples stored at
Rheumatoid Factor 200 IU/mL
Turbidimetric method. room temperature for 24 hours retain 80 percent of their initial 567 001 50 test
concentration. Stored at -20°C, it is stable for 6 months. 3 cycles of
Freeze/thaw has <2% loss of PCT in sample. Analytical characteristics
Test Principle
PCT Assay is based on a latex enhanced immunoturbidimetric assay.
PCT proteins in the sample bind to the specific anti-PCT antibody, Stability: several days at 4oC Precision
which is coated on latex particles, and causes agglutination. The 24 hours at room temp. Within-run precision of the Diazyme PCT Assay was evaluated. In
degree of the turbidity caused by agglutination can be measured 6 months at -20 oC the study, three levels of PCT controls containing 0.5 ng/mL,
optically and is proportional to the amount of PCT in the sample. 2.14 ng/mL and 13.13 ng/mL PCT respectively were tested with 20
The instrument calculates the PCT concentration of a sample by System Parameters replicates in one run.
interpolation of the obtained signal of a 6-point calibration curve Wavelength 600 nm Level 1 Level 2 Level 3
Optical path 1 cm
Assay type Turbidimetric Number of Data Points 20 20 20
Reagents
Reagent 1 (R1 Buffer) Sample: Reagent Ratio 1 : 60 Mean (ng/mL) 0.50 2.14 13.13
Tris- buffer 100 mmol/L Temperature 37 oC and 20 – 25 oC
SD (ng/mL) 0.036 0.069 0.353
Reagent 2 (R2) Zero adjustment Reagent or Sample blank
Suspension of anti-human PCT mouse monoclonal antibody Linearity 60 ng/mL CV (%) 7.2% 3.2% 2.7%
coated latex particles (0.2%), ready to use
Procedure Accuracy
Calibrator : contain PCT in a human serum matrix lyophilized The performance of this assay was compared with the performance
Calibrator Sample
material. of a legally marketed PCT Assay using lithium heparin plasma
R1(buffer) 270 ml 270 ml samples. For the 41 plasma samples with PCT ranging from 0.18
Materials required but not provided with the kit Sample ------ 25 ml ng/mL to 58.44 ng/mL, the correlation coefficient between the two
Any instrument with temperature control of 37 ± 0.5°C that is methods was 0.9141, slope was 0. 9815, and y intercept was 0.3137
Calibrator 25 ml ------
capable of reading absorbance accurately at 600 nm may be used.A
saline solution is needed for calibrator and high sample dilution.The Mix and incubate for 4.5 minutes at 37 C,then add o Limit of Quantitation
PCT Calibrator Set and PCT Control Set are sold separately The Limit of Quantitation (LOQ) of the PCT Assay was determined
R2 Latex 90 ml 90 ml
to be 0.17 ng/mL..
Mix and incubate for 2 minutes,measure absorbance of
Precautions and Warnings Expected Values
specimenA1,then after 3 minutes record absorbance of specimenA2
1. For “In Vitro Diagnosticc Use” 1.Spectrum Procalcitonin < 0.5 ng/mL: systemic infection (sepsis)
2. Do not use the Reagent after the expiration date labeled on is not likely, Local bacterial infection is possible; PCT > 0.5 ng/mL
Calculation and < 2 ng/mL: systemic infection (sepsis) is possible but other
the outer box.
Generate a reference curve by successive 1 : 2 dilutions of standard
3. Assay calibration frequency is dependent on instrument used. conditions are known to increase PCT as well; PCT > 2 ng/mL and
Additionally, the assay should be recalibrated and controls < 10 ng/mL: systemic infection (sepsis) is likely, unless other cause
run with each new lot of reagents. are known; PCT > 10 ng/mL, important systemic inflammatory
following:
4. Avoid ingestion and contact with skin and eyes. response, almost exclusively due to severe bacterial or septic shock.
D absorbance of sample = (A2 - A1) sample
5. Specimens containing human sourced materials
D absorbance of each standard = (A2 - A1) for each standard
should be handled as if potentially infectious using safe
Plot the calibration curve and obtain the result.
laboratory procedures, such as those outlined in Biosafety in
Microbiological and Biomedical Laboratories.
6. The reagent contains < 0.1% sodium azide, NaN3, as
preservative. Sodium azide may react with lead and copper
plumbing to form highly explosive metal azide. On disposal,
flush with a large volume of water to prevent azide buildup.
224 225
RF Control SYMBOLS IN PRODUCT LABELLING RF Standard Super High SYMBOLS IN PRODUCT LABELLING
REF: 344 001 1x1 ml REF 349 001 1x1 ml
Specifications
Intended Use
Accuracy control for the determination of Rheumatoid factor (RF) in serum by turbidimetry and nephelometry.
Lot number : RF 1014
composition
A dilution of humen plasma containing high levels of RF with saline. Expiration Date : October 2016
The dilution is liquid stabilized and contains : 0.095 g% sodium azide as preservative.
Source : Human plasma
Storage and Stability o
The expiry date of the product at +2 to +8 ºC is listed on the label. Storage: 2-8 C Do Not Freeze
After first opening the control can be used for 6 weeks if stored tightly closed
at +2 to+8 ºC after use. Stabilitty after opening the vial : 6 weeks
Do not freeze.

1. For “In Vitro Diagnosticc Use”
2. Each individual donation intented for use in manufacturing of protein control serum was tested for hepatities B surface antigin
(HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were
used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in
accordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven .
Assigned Value
LOT No.: RFT 1014
RF : 103 ( 88 - 118 ) IU/mL.
Target RF IU/mL
RF Standard Super High
498
226 227
RHEUMATOID FACTOR (RF) SYMBOLS IN PRODUCT LABELLING
Calculation
Generate a reference curve by successive 1 : 2 dilutions of calibrator
ImmunoTurbidimetry EC REP Authorised Representative
3rd Generation LOT Batch Code/Lot number
Catalogue Number
for use
as following:
D absorbance of sample = (A2 - A1) sample
(Aggregated human IgG method) REF
D absorbance of each calibrator = (A2 - A1) for each calibrator
Plot the calibration curve and obtain the result..
REF: 598 001 100 test
R1 Buffer 2 x 20 ml Senstivity
3 IU /mL.
R2 Reagent 2 1 x 8.2 ml
Linearity
Intended Use As with other diagnostic tests, results should be interpreted 500 IU /mL.
In vitro diagnostic reagents for the quantitative determination of considering
Rheumatoid Factor (RF) in human serum by means of particle- all other test results and the clinical situation of the patient. Quality Control
enhanced turbidimetric immunoassay. Control sera are recommended to monitor the perfomance of manual
Reagent Preparation, Storage and Stability and automated assay procedures .
Background All reagents are supplied ready to use. Each laboratory should establish its own Quality Control Scheme and
The most consistent serological feature of rheumatoid arthritis is Reagents in the original vial are stable to the expiration date on the corrective actions if controls do not meet the acceptable tolerances.
the increased concentration of autoantibodies directed against vial label when capped and stored at (2 - 8 ºC).
antigenic sites in the Fc region of human and animal IgG, namely Expected Values
rheumatoid factors (RFs) in the blood and joint fluid. The potential RF Standard: 0-20 IU/mL.
role of these factors in the pathogenesis of this disease has been The Standard is stable to the expiration date on the vial label when Each laboratory should establish an expected range for the
studied extensively, with the finding that both environmental and capped and stored at (2 - 8 ºC). geographical area in which it is located..
genetic factors affect production of RF. RF determinations are Once opened the Standard is stable for 6 weeks if stored tightly
clinically important for the diagnosis, prognosis, and assessment of closed at 2 - 8 ºC after use. References
therapeutic efficacy of rheumatoid arthritis. The RF is a term used to 1. Arnet FC, Edworthy SM, Bloch DA, McShane DJ, et al. The
describe a variety of antibodies (in most cases of the IgM type) that For further storage at - 30 ºC divide Standard into aliquots. Stability American Rheumatism Association 1987. Revised criteria for
will react with modified human IgG (e.g. IgG in circulating immune 3 months. once thawed never freeze again. the classification of rheumatoid arthritis. Arthritis Rheum 1988;
complexes, IgG adsorbed to latex, etc.) and IgG of animal origin. 31:315-24.
RF is highly associated with rheumatoid arthritis, as high as 90 % of Specimen collection and preparation 2. Bandilla,K.I. and MC DUffie,F.C,Arthritis Rheum,12,74(1969).
patients with RA have RF titers of more than 20 IU/mL. Fresh serum . stable 2 days at2 - 8 ºC or 3 months at - 20 ºC the 3. Bartfield H. Distribution of rheumatoid factor activity in non
samples with presence of fibrin should be centrifuged before rheumatoid states. Ann NY Acad Sci 1969; 168:30-40. Singer
Test Principle testing Do not use highly hemolized or lipemic samples . JM, Plotz CM. The latex fixation test. Am J Med 1956; 21:888-
This RF test is based upon the RF antigen-antibody reaction 92.
Procedure 4. Muller,W,The Serology of Rheumatoid Arthritis.Berlin-
Reagents 1. Bring the reagents and the photometer to 37ºC Gottingen-Heidelberg 97(1962).
2. Assay conditions: 5. Moore TL, Dorner RN. Rheumatoid factors. Clin Biochem 1993;
R1 Buffer Wavelength 340 nm 26:75- 84.
50 mmol/L Good’s buffer (pH7.4). Temperature 37ºC 6. Waaler,e,Acta Path.Microb.scan,17 (1940).
Sodium azide (0.95 g/L). Cuvette 1cm light path
3. Adjust the instrument to zero with distilled water .
R2 reagent 2 4. Pipette into a cuvette ORDERING INFORMATION
Heat-aggregated human IgG(<0.5 mg/ml). CATALOG NO QUANTITY
Sodium azide (0.95 g/L) Standard Sample
598 001 100 test
Standard 25 ml ----
1- Standard
Sample ---- 25 ml
RF concentration is stated on the vial label.
Mix and incubate for 2 minutes,read absorbance (A1)
2-Controls
For in vitro diagnostic use only. Do not pipette by mouth. Reagents
containing sodium azide must be handled with precaution. Sodium After addition of R2, incubate and after 5 minutes record 2nd reading
azide can form explosive azides with lead and copper plumbing. (A2)
and reagents obtained from human blood should always be handled
with precaution using established good laboratory practices.
Disposal of all waste material should be in accordance with local

guidelines.
228 229
RHEUMATOID FACTOR (RF) SYMBOLS IN PRODUCT LABELLING Blank Sample / Calibrator ORDERING INFORMATION

NaCl 9 g/L
Sample / Calibrator
4 ml
----
----
4 ml
CATALOG NO QUANTITY
561 001 100 test

REF Catalogue Number for use 7. Mix and read the absorbance after 2 minutes (A2) of the
REF: 561 001 100 test sample addition.
R1 Buffer 2 x 20 ml Calculation
R2 Latex 1 x 10 ml Calculate the absorbance difference (A2-A blank) of each point of
preparation the calibrator dilution and plot the values obtained against the RF
Intended Use RF Calibrator : Reconstitute with 2 ml of distilled water : Mix gentity concentration of each calibrator dilution .
In vitro diagnostic reagents for the quantitative determination of and bring to room temperature for about 10 minutes before use Rheumatiod factor concentration in the sample is calculated by
Rheumatoid Factor (RF) in human serum by means of particle- Calibration Curve : interpolation of its (A2-Ablank) in the calibration curve.
prepare the following RF Calibrator dilutions in NaCl 9 g/L
enhanced turbidimetric immunoassay.
Muttiply the concentration of the RF Calibrator by the corresponding
factor stated in table below to obtain the RF concentration of each Quality Control
Background dilution. Control sera are recommended to monitor the perfomance of manual
The most consistent serological feature of rheumatoid arthritis is Calibrator dilution 1 2 3 4 5 6 and automated assay procedures .
the increased concentration of autoantibodies directed against Calibrator RF --- 25 50 100 200 400 Each laboratory should establish its own Quality Control Scheme
antigenic sites in the Fc region of human and animal IgG, namely Na Cl 9 g/L 400 375 350 300 200 ---
and corrective actions if controls do not meet the acceptable
rheumatoid factors (RFs) in the blood and joint fluid. The potential tolerances
Factor 0 0.0625 0.125 0.25 0.5 1
role of these factors in the pathogenesis of this disease has been
Concentration(IU/ml)
studied extensively, with the finding that both environmental and (for example: Expected Values
0 12.5 25 50 100 200
genetic factors affect production of RF. RF determinations are the undiluted C = Up to 20 IU/mL.
200 IU/ml )
clinically important for the diagnosis, prognosis, and assessment Each laboratory should establish its own reference range.
of therapeutic efficacy of rheumatoid arthritis. Although RFs may
be found in all immunoglobulin classes, the RF most frequently Storage and Stability Senstivity
detected in the laboratory is IgM type, present in about 75 - 80 % of All the compontents of the kit are stable until the expiration date on 6 IU /mL.
adult patients with rheumatoid arthritis but in about 10 % of children the label when stored tightly closed at (2 - 8 ºC) and contaminations
with juvenile rheumatoid arthritis. prevented during their use .Do not use reagents over the expiration Linearity
date . 160 IU /mL.
Test Principle Reagnt deterioration : presence of paticles and turbidity specimens showing higher concentration should be diluted 1+4
This RF test is based upon the reactions between IgM-anti-IgG (RF) Reconstituted calbibrator :stable for 1 month at 2 - 8 ºC or 3 using physiological saline and repeat the assay (result×5).
in patient’s sample and latexcovalently bound human IgG. RF values months at 20 ºC
are determined photometrically. interferences
Do not freeze: frozen Latex or Diluent couled change the Hemoglobin (10 g/L) , bilirrubin (20 mg/dL) and lipemia (10 g/L) ,
Reagents functoinality of the test. do not interfere.Other substances may interfere6.
Buffer Samples References

Phosphate buffer (0,05 M) pH: 8,0 containing NaCl (0,15M), Fresh serum or plasma . stable 7 days at2 - 8 ºC or 3 months at 20 1. Arnet FC, Edworthy SM, Bloch DA, McShane DJ, et al. The
detergent and polyethyleneglycol. ºC the samples with presence of fibrin should be centrifuged before American Rheumatism Association 1987. Revised criteria for
Preservative : sodium azide < 1g/L testing Do not use highly hemolized or lipemic samples . the classification of rheumatoid arthritis. Arthritis Rheum 1988;
31:315-24.
Latex reagent Procedure 2. Bartfield H. Distribution of rheumatoid factor activity in non
A suspension of latex microparticules covalently bound human IgG 1. Bring the reagents and the photometer to 37ºC rheumatoid states. Ann NY Acad Sci 1969; 168:30-40. Singer
in a glycin buffer (0,1 M, pH: 8,2), containing NaCL 2. Assay conditions: JM, Plotz CM. The latex fixation test. Am J Med 1956; 21:888-
(0,15 M) and bovine serum albumin (0,5%). Wavelength 650 nm (600 -650 nm) 92.
Preservative: Sodium azide 0,075% Temperature 37ºC 3. Moore TL, Dorner RN. Rheumatoid factors. Clin Biochem 1993;
Cuvette 1cm light path 26:75- 84.
Calibrator 3. Adjust the instrument to zero with distilled water . 4. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th
Human - based reference fluid. Preservative: sodium azide, 0.075 4. Pipette into a cuvette : Edition, AACC Press, 2000.
%. blank 5. Passing H, Bablok W. A new biometrical procedure for testing
All raw materials of human origin used in the manufacture of this Diluent (ml) 0.4 the equality of measurements from two analytical methods.
product showed no reactivity when tested for HBsAg, anti-HIV-1/2 6. Application of linear regression procedures for method
Latex (ml) 0.1
and HCV with commercially available test methods. comparison studies. Part I. J Clin Chem Clin Biochem 1983;
However, this product should be handled as though capable of 5. Mix and read the absorbance (blank reagent) 21:709-20.
transmitting infectious diseases 6. add the sample / Calibrator 7. Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26:
207 – 224
230 231
Specific Protein Control SYMBOLS IN PRODUCT LABELLING Specific Protein standard SYMBOLS IN PRODUCT LABELLING
REF:390 001 1x1 ml REF:387 001 1x1 ml
Intended Use Intended Use

Accuracy control for the determination of protiens in serum by turbidimetry and nephelometry. Accuracy control for the determination of protiens in serum by turbidimetry and nephelometry.
composition composition
A dilution of human plasma with phosphsate buffered saline. A dilution of humen plasma with phosphsate buffered saline.
Contains stabilizers and 0.095 g% sodium azide . Contains stabilizers and 0.095 g% sodium azide .
Storage and Stability Storage and Stability

The expiry date of the product at +2 to +8 ºC is listed on the label. The expiry date of the product at +2 to +8 ºC is listed on the label.
After first opening the control can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use. After first opening the standard can be used for 6 weeks if stored tightly closed at +2 to+8 ºC after use.
Do not freeze. Do not freeze.
Precautions and Warnings Precautions and Warnings

1. For “In Vitro Diagnostic Use” 1. For “In Vitro Diagnostic Use”
2. Each individual donation intended for use in manufacturing of protein control serum was tested for hepatities B surface antigin 2. Each individual donation intended for use in manufacturing of protein control serum was tested for hepatities B surface antigin
(HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were (HBsAg),anti-hepatitis C virus (anti-HCV) and anti-HIV1 and HIV2 by FDA required test. Only donations with negative findings were
used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in used for its manufacture.Nevertheless every product obtained from human body fluids should be handled with appropriate care in
accordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven . accordance with recommended procedures for biohazardous materials since absence of infectious agents can never be proven .
3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes ! 3. Reagents containing sodium azide must be handled with due caution: Do not ingest or allow to contact skin or mucous membranes !
Lot No.: MPP011

Lot No.: PED011
Protein Values
Protein Values (mg/dl) Protein
(mg/dl)
Protein
Target Range
a2-Macroglobulin 33.3
a2-Macroglobulin 11.1 9.4 - 12.8

Transferrin 39.9
Transferrin 13.3 11.3 - 15.3

IgA 28.7
IgA 9.6 8.1 - 11.0

IgG 131.6
IgG 43.9 37.3 - 50.4

IgM 18.0
IgM 6.0 5.1 - 6.9

Kappa Light chain* 32.7
Kappa Light chain* 10.9 9.3 - 12.5

Lambada light chain* 19.1
Lambada light chain* 6.4 5.4 - 7.3

Values based on a reference preparation (ERM-DA470K/IFCC) from the International Federation of clinical chemistry (IFCC).
Values based on a reference preparation (ERM-DA470K/IFCC) from the International Federation of clinical chemistry (IFCC).
232 233
E-Infectious
i m m u n o l o g y,
Bacteriology
234 235
Brucella A Brucella M SYMBOLS IN PRODUCT LABELLING Slide Semi-quantitative Method Remarks
EC REP Authorised Representative Temperature Limitation 1. Using a pipette place 80ml, 40ml, 20ml, 10ml and 5ml of patient 1. Turbid and contaminated serum should not be used for testing
IVD For in-vitro diagnostic use Use by/Expiration Date serum to be tested on 5 different reaction circles on the glass 2. In the semi quantitative test the reactions obtained are roughly
REF: 710 001 50 test REF: 710 003 50 test slide. The corresponding titres obtained will be 1:20, 1:40, 1:80, equivalent to those which would occur in tube test.
REF: 710 002 100 test REF: 710 004 100 test 1:160 & 1:320 respectively. 3. Agglutinins are found in high proportion of normal individuals
2. Place one drop of appropriate Brucella antigen suspension to and titres less than 1:80 are of doubtful significance. A rising
each circle. titre is more significant than a single
Brucella Combo 3. Gently Rock the slide back and forth, and observe for high titre
agglutination macroscopically at one minute against a white 4. False positive reactions may occur in sera of patients infected
background. with Pasteurella tularensis or vaccinated with vibrio Cholerae.
REF: 710 005 50 test REF: 710 005 -1 100 test 5. False positive results are likely if the test is read more than one
Tube-test procedure minute after mixing on slide test.
1. Take 8 test tubes and label them 1 to 8. 6. Its recommended that results of the tests should be correlated
Intended Use Specimen Collection and Storage 2. Pipette 1.9 ml of isotonic saline or preferably 0.25% phenol with clinical findings to reach the final diagnosis.
Spectrum Diagnostics Brucella reagent is intended for the detection 1. No special preparation of the patient is required prior to sample saline to tube No.1. 7. Prozoning may sometimes be encountered in serum containing
of Anti- Brucella antibodies in human serum. collection by approved techniques. Do not use haemolysed 3. To each of the remaining tubes (2-7) add 1.0 ml of isotonic very high titres on slide test.
samples. saline or preferably 0.25% phenol saline 8. Since techniques and standardization vary from lab to lab one
Background 2. Clean and dry glassware free from detergents must be used for 4. To the tube No. 1 add 0.1 of serum sample to be tested and tube difference in tube titres can be expected.
Human Brucellosis (Diurnal or undulant fever) is a common febrile sample collection. mix well.
illness caused by infection with bacteria of some of the Brucella 3. Don’t heat inactive the serum. 5. Transfer 1 ml of the diluted serum sample from tube No. 1 to Warranty
species (Abortus, Melitensis). This undulant fever is associated with 4. Freshly collected serum is preferable, store samples at tube No 2 and mix well. This product is designed to perform as described on the label and
o
symptoms, which are often variable and non-specific with chills, 5. 2 – 8 C for 24 hours or frozen for several days. 6. Transfer 1 ml of the diluted serum sample from tube No. 2 to package insert, the manufacturer disclaims any implied warranty of
fever, sweats and anorexia. On exposure the body responds to this tube No. 3 and mix well. Continue this serial dilution till tube use and sale for any other purpose.
antigenic stimulation by producing specific antibodies whose titres Materials provided with the kit No. 7 in each set.
rise slowly at early stages and then increases. Specific antibodies Stained Brucella-A / Brucella-M antigen suspension 7. Discard 1.0 ml of the diluted serum from tube No. 7. References
to the Brucella species are detectable a few weeks after exposure 8. Pipette 1.0 ml of isotonic saline in tube No. 8, which serves as 1. J. G. Collee, J.P. Duguid, A G Fraser. Practical Medical
and are of considerable importance in the diagnosis of Brucellosis. Additional material required negative control. Microbiology, 13 th Ed: 525-530.
Information regarding the titre of antibodies can be obtained by 9. To all the tubes add one drop of appropriate Spectrum Brucella 2. G.Galton, L.M.Jones, R.D.Angus, J.M.Verger. techniques for
using specific Spectrum Brucella antigen suspensions. Slide test method: antigen suspensions and mix well the Brucellosis laboratory,© INRA, Paris, 1988.
Stop watch, positive control, isotonic saline, and glass slide with 10. Cover and incubate at 37oC for 24 hours. 3. Felix A., (1942), Brit. Med. J., 11, 597.
Assay Principle clear/white background, appropriate pipettes/Micropipettes, mixing 11. Observe for agglutination macroscopically in each tube of the
The smooth, killed stained Brucella antigen suspensions are mixed sticks & a high intensity direct light source. dilution series.
with the patient’s serum. Specific antibodies to Brucella antigens if ORDERING INFORMATION
present in the patient serum will react with the antigen suspension Quantitative method: Interpretation of the results CATALOG NO QUANTITY
to produce an agglutination reaction. No agglutination indicates the Timer, test tube (12 mm x 75 mm), test tube rack, appropriate
Brucella A (Abortus)
absence of specific antibodies to Brucella antigens. pipettes/Micropipettes, isotonic saline/0.25% phenol saline and Slide screen method
o
Incubator (37 C). Agglutination is a positive test result and indicates the presence 710 001 2.5 ml 50 test
Reagent of specific antibodies to Brucella in the patient serum. 710 002 5 ml 100 test
Spectrum Brucella-A / Brucella-M reagents contain ready to use Procedure No agglutination is a negative test result and indicates absence of Brucella M (Melitensis)
standardized, killed, stained, smooth specific antigen suspensions (a) Bring reagents to room temperature. specific antibodies to Brucella in the patient serum. 710 003 2.5 ml 50 test
of Brucella having specific reactivity towards antibodies to Brucella (b) Shake and mix Brucella antigen suspensions well before 710 004 5 ml 100 test
abortus (Brucella-A) and Brucella melitensis (Brucella-M). dispensing. Slide Semi-Quantitative method
Brucella Combo
(c) The procedure for Brucella-A / Brucella-M is indentical. Agglutination is a positive test result. The titre of the patient serum
Reagent Storage and Stability 2.5 ml Brucella A Reagent
corresponds to the visible agglutination in the test circle with the
710 005 2.5 ml Brucella M Reagent
1.
o
Store the reagents at 2 – 8 C (Do not freeze). Slide test Method minimum amount of serum sample.
0.7 ml Positve control
2. The shelf life of reagent is as per the expiry date mentioned 1. Place one drop of positive control onto a reaction circle of the
5 ml Brucella A Reagent
on the reagent vial labels. Each batch of reagents undergoes glass slide. Tube-test method
710 005-1 5 ml Brucella M Reagent
rigorous quality control at various stages of manufacture for its 2. Place 80ml of saline onto the next reaction circle of the glass The titre of the patient serum is the last dilution of the serum sample
0.7 ml Positve control
specificity, sensitivity and performance. slide. that gives a granular agglutination.
3. Place 80ml of patient serum to be tested onto each of the In negative reaction, the appearance of the suspension remains
Note required number of reaction circle. unchanged, which show a typical swirl when the tube is flicked
1. In-vitro diagnostic reagent for laboratory and professional use 4. Add one drop of the appropriate Spectrum Brucella antigen
only. Not for medicinal use. suspension in each of the above circles. (containing positive
2. The reagent contains 0.01% thimerosal as preservative. Avoid control & saline and the patient serum to be tested).
contact with skin and mucosa. On disposal flush with plenty of 5. Mix contents of each circle uniformly over the entire circle with
water. separate mixing sticks.
6. Gently Rock the slide back and forth, and observe for
agglutination macroscopically at one minute against a white
background.
236 237
Widal Test SYMBOLS IN PRODUCT LABELLING Slide Semi-quantitative Method Remarks
EC REP Authorised Representative Temperature Limitation 1. Using a pipette place 80µl, 40µl, 20µl, 10µl and 5µl of patient 1. Positive results obtained in the slide test should be confirmed
(Salmonella Ab) IVD
LOT
serum to be tested on 5 different reaction circles on the glass with the tube test to establish whether the titres are
slide. The corresponding titres obtained will be 1:20, 1:40, 1:80, diagnostically significant or not.
1:160 & 1:320 respectively. 2. TAB vaccinated patients may show a high titre of antibodies to
2. Follow step No. 5 -7 of slide screen method. each of the antigens.
REF: 718 001 50 test 4 x 2.5 ml 3. “O” being a somatic antigen brings about a coarse, compact,
REF: 718 002 100 test 4 x 5 ml Note: granular agglutination whereas “H” being a flagellar antigen
This method is recommended for obtaining quick approximate titres brings about larger, loose, flocculant agglutination.
only. 4. While the “O” antigen is species specific, the”H” antigen is
Intended Use Specimen Collection and Storage specific to the serotype.
Spectrum Diagnostics Widal set is intended for the detection of anti- 1. No special preparation of the patient is required prior to sample Quantitative Method: 5. Turbid and contaminated sera should not be used for testing.
salmonella antibodies in human serum. collection by approved techniques. Do not use haemolysed 6. Generally antibody titres of 1:80 or more are considered
samples. Tube-test procedure clinically and diagnostically significant. However the significant
Background 2. Clean and dry glassware free from detergents must be used for 1. Take appropriate number of sets (as required: one set for each titre may vary from population to population and needs to
Ecteric fever occurs when pathogenic microorganisms like S.typhi, sample collection. antigen suspension) of 8 kahn tubes / test tubes and label them be established for each area.
S.paratyphi A, S.paratyphi B, S.paratyphi C infect the human body. 3. Don’t heat inactivate the serum. 1 to 8. 7. Its recommended that results of the tests should be correlated
o
During the course of disease, the body responds to this antigenic 4. Freshly collected serum is a preferable, store sample at 2 – 8 C 2. Pipette into tube No. 1 of all sets 1.9 ml of isotonic saline with clinical findings to arrive at the final diagnosis.
stimulus by producing antibodies whose titre rise slowly in early in case of delay in testing, for up to 72 hours. 3. To each of the remaining tubes (2 to 8) add 1 ml of isotonic 8. Since techniques and standardization vary from lab to lab one
stages, to a maximum and then slowly falls till it is undetectable. saline tube difference in tube titres can be expected.
Antibodies to salmonella organisms may be detected in the patient Material Provided with the Kit 4. To tube No. 1 of all sets add 0.1 of serum sample to be tested 9. The performance of the reagents should be validated
serum from the second week after onset of infection. Information 1. S.typhi “O” Antigen suspension. and mix well. occasionally using the positive control provided. Good
regarding the titres and whether or not they are rising or falling can be 2. S.typhi “H” Antigen suspension. 5. Transfer 1 ml of the diluted serum sample from tube No. 1 to physiological saline may be used as a negative control
obtained by performing serological tests using Spectrum salmonella 3. S.paratyphi “AH” Antigen suspension. tube No 2 and mix well.
antigen suspensions. Usually tube titres of 1:80 and above are taken 4. S.paratyphi “BH” Antigen suspension. 6. Transfer 1 ml of the diluted serum sample from tube No. 2 to References
as diagnostically significant, however for endemic areas higher 5. Polyspecific positive control (Goat). tube No. 3 and mix well. Continue this serial dilution till tube No. 1. Cruickshank R., (1982), Medical Microbiology, 12th Edition,403.
cut-offs may need to be established. 7 in each set. 2. Felix A., (1942), Brit. Med. J., 11, 597.
Note: item Nos. 1- 4 each is available as individual reagent pack. 7. Discard 1.0 ml of the diluted serum from tube No. 7 of each set
Assay Principle 8. Now the dilutions of the serum sample achieved from tube No.
When the colored, smooth, killed salmonella antigen suspensions Additional material required 1 to 7 respectively in each set is as follows 1:20, 1:40, 1:80, ORDERING INFORMATION
are mixed/incubated with patient serum, anti-salmonella antibody Slide test method: stop watch, variable micropipettes and mixing 1:160 , 1:320 1:640, 1:1280. tube no 8 in all the sets, serves as CATALOG NO QUANTITY
present in the patient serum react with the antigen suspensions to sticks. a saline control.
718 001 4 x 2.5 ml 50 test
give agglutination. Agglutination is a positive test result, indicating Quantitative method: Time, Kahn Tubes / test tubes, pipettes 9. To all the tubes (1 to 8) of each set add one drop of all 718 002 4 x 5 ml 100 test
o
presence of anti-salmonella antibodies in the patient serum. No (0.1 ml, 1 ml), isotonic saline, incubator (37 C), test tube rack. respective well mixed Spectrum Salmonella antigen
agglutination is a negative test result indicating absence of anti- suspensions from the reagent vials and mix well
salmonella antibodies. Procedure o
10. Cover and incubate at 37 C overnight (approximately 18 hours)
(a) Bring reagents to room temperature before testing. 11. Dislodge the sedimented button gently and observe for
Reagents (b) Shake and mix antigens well before dispensing. agglutination.
Spectrum widal kit contains ready to use concentrated, smooth
antigen suspensions of the bacilli; S.typhi “O”, S.typhi “H”, Slide Screen Method Interpretation of the results
S.paratyphi “AH”, S.paratyphi “BH” and polyspecific positive control 1. Place one drop of positive control onto a reaction circle of the
reactive with these antigens. Each batch of reagents undergoes glass slide. Slide screen method
rigorous quality control at various stages of manufacture for its 2. Place one drop (50 ml) of isotonic saline onto the next reaction • Agglutination is a positive test result and indicates presence of
specificity and performance. circle of the glass slide. the corresponding antibody in the patient serum.
3. Place one drop (50 ml) of patient serum to be tested onto each • No agglutination is a negative test result and indicates absence
Reagent Storage and Stability of the required number of reaction circle. of the corresponding antibody in the patient serum.
o
1. Store the reagents at 2 – 8 C (Do not freeze). 4. Add one drop of appropriate Spectrum Salmonella antigen
2. the shelf life of reagent is as per the expiry date mentioned on suspension to the reaction circles containing positive control Slide Semi-Quantitative method
the reagent vial labels. & isotonic saline. Agglutination is a positive test result. The titre of the patient serum
5. Add one drop of appropriate Spectrum Salmonella antigen corresponds to the visible agglutination in the test circle with the
Note suspensions to the reaction circles containing the patient smallest amount of serum sample.
1. In vitro diagnostic reagent for laboratory and professional use serum.
only. Not for medicinal use. 6. Mix contents of each circle uniformly over the entire circle with Quantitative method
2. The S. typhi “O” reagent contains 0.5% phenol, S.typhi “H”. S. separate mixing sticks. The titre of the patient serum using Spectrum Salmonella antigen
paratyphi “AH”, and S. paratyphi “BH” reagents contain 0.3% 7. Rock the slide gently back and forth, and observe for suspensions is the highest dilution of the serum sample that gives a
formaldehyde as preservatives. Avoid contact with skin and agglutination macroscopically at one minute. visible agglutination.
mucosa. On disposal flush with large quantities of water.
3. Only a clean and dry glass slide must be used. Clean the glass
slide with distilled water and wipe dry.
238 239
RPR SYPHILIS TEST QUALITATIVE PROCEDURES PROCEDURE LIMITATION
1. Bring reagents to room temperature. 1. This test provides a presumptive diagnosis of syphilis.
2. Dispense 50ul of sample onto a single circle on the test card. Physicians should evaluate all clinical and laboratory findings
REF 725 001 100 test 3. Repeat step 2 for the positive and negative controls. before making a definitive diagnosis.
REF 725 002 200 test 4. Spread the sample of each test specimen over the entire test 2. In positive specimens, it is recommended to confirm the result
REF 725 000 100 test (Carbon Antigen Reagent Only) circle. by another serological test such as the TPHA.
5. Mix the carbon antigen suspension well.
6. With the needle suck up reagent sufficient to the testing REFERENCES
A qualitative and quantitative test for the detection of Non-Treponema requirements. 1. McGrew B.E., Stout G.W., Falcon V.H., AM. J. Med. Techs.,
in serum or plasma 7. Dispense one free-fall drop of the carbon antigen onto each 34:634, 1969
test circle containing specimen. Do not mix the antigen with 2. Manual of Tests for Syphilis, PHS publication No.411, 1969.
INTRODUCTION & PRINCIPLE the sample. 3. Larsen S.A., et. al., ata on file, Treponemal Research and
Besides other antibodies, Treponema Pallidium produces non- 8. Using the rotator, rotate the card at 100 rpm for 8 minutes. Immunology lab, CDC.
Treponemal antibodies (regain) in syphilitic persons. These 9. Read the results in good light immediately after 8 minutes.
antibodies can be detected by RPR antigen. SPECTRUM RPR
card test is a macroscopic screening test for the qualitative and READING THE QUALITATIVE RESULTS
quantitative detection of reagin antibodies in serum or plasma. POSITIVE - If large aggregates appear in the centre or the periphery ORDERING INFORMATION
The kit contains RPR antigen which is based on the easy to use of the test circle containing the sample. If the aggregates are visible, CATALOG NO QUANTITY
VDRL carbon antigens. In the presence of the reagin, the antigen but weak or small, then the test should be read as
725 001 100 test
causes flocculation of the carbon particles, which appears as black WEAK POSITIVE. If test is positive, then results should be confirmed 725 002 200 test
clumps. The charcoal particles contained in the antigen suspension by the quantitative procedure mentioned below. 725 000 100 test (Carbon Antigen Reagent Only)
enhances the visual appearance of the coagglutination in positive NEGATIVE - If no aggregates appear and the specimen has smooth
samples. grey appearance.
PREPARING THE SPECIMEN QUANTITATIVE PROCEDURES

SPECTRUM RPR kit can be used with either unheated plasma or 1. Dispense 50ul of 0.9 % saline onto to test circles numbered 2
heated serum samples. Serum samples can stay stable for up to 5 to 5. Saline should not be spread. Dispense 50ul of specimen
o
days if stored at 2 to 8 C. Plasma samples collected with EDTA can onto test circle 1.
o
stay stable up to 24 hours if stored at 2 to 8 C. 2. Dispense 50ul of specimen onto test circle 2. Prepare serial
two-fold dilutions by drawing the mixture up and down the
MATERIALS PROVIDED pipette 5-6 times ( avoid any bubble formation. Transfer 50ul
1. SPECTRUM RPR carbon antigen reagent from circle 2 to 3, to 4 and to 5. Dispose 50ul from circle 5 after
2. Positive controls mixing.
3. Negative controls 3. Starting from circle 5 and onto 4,3,2 and 1, mix and spread the
4. RPR tests cards serum over the entire area of each test circle.
5. 20 G dispensing needle (16 ul/drop) 4. Continue with steps 6 - 9 of the qualitative procedure.
MATERIALS NEEDED BUT NOT PROVIDED READING THE QUANTITATIVE RESULTS

Saline 0.9 %, rotator (100rpm), accurate pipette to deliver 50ul and The dilutions of the circles are as follows:
timer. Circle 1 2 3 4 5
Dilution - 1:2 1:4 1:8 1:16
PRECAUTIONS
The titer of the sample is read as follows (P: Positive, N: Negative)
1. The reagents in this kit should be stored in an upright position
Positive 1:2 P P N N N
and refrigerated between 2 to 8. Never Freeze. Test cards need
not to be refrigerated and can be kept at room temperature. Positive 1:4 P P P N N
2. Reagents should be brought to room temperature and mixed Positive 1:8 P P P P N
well to obtain a uniform suspension of carbon particles. Positive 1:16 P P P P P
3. After use, dispensing Dropper and needle should be washed
well with distilled water then air dried. Positive and negative results are read as in the reading qualitative
4. Stability of the antigen may be reduced if stored in the plastic results procedure.
dispensing Dropper for a long time. It is highly recommended If the result in circle 5 is positive, then further dilution to 1:32, 1:64,
to return the antigen to the original glass Dropper at the end of 1:128 and 1:256 is required. Use steps 3 in quantitative procedure
the testing session. and steps 6-10 in qualitative procedure to obtain the required
5. Always use a fresh pipette tip for every test. dilutions.
240 241
Rose Bengal Brucella Antigen SYMBOLS IN PRODUCT LABELLING
REF: 711 001 50 test
REF: 711 002 100 test
REF: 711 003 100 test + Positive Control LOT Batch Code/Lot number CAUTION. Consult instructions
Intended Use Additional material required

Spectrum Diagnostics Brucella Rose Bengal reagent is intended Slide test method:
for the early detection of Brucella specific agglutinins (Brucella Stop watch, positive control, isotonic saline, and glass slide
Abortus, Melitensis and Suis). with clear/white background, appropriate pipettes/Micropipettes,
mixing
Background sticks & a high intensity direct light source.
Human Brucellosis (Diurnal or undulant fever) is a common
febrile illness caused by infection with bacteria of some of the Procedure
Brucella species (Abortus, Melitensis). This undulant fever is 1. Allow reagents and serum samples to reach room
associated with symptoms, which are often variable and non- temperature before use.
specific with chills, fever, sweats and anorexia. On exposure 2. Shake the antigen bottle gently to insure a uniform
the body responds to this antigenic stimulation by producing suspension.
specific antibodies whose titres rise slowly at early stages and 3. Place one drop (40ml) serum sample to the selected ring of
then increases. Specific antibodies to the Brucella species are the slide.
detectable a few weeks after exposure and are of considerable 4. Place one drop of the Rose Bengal antigen to serum sample.
importance in the diagnosis of Brucellosis. Information regarding 5. Mix serum sample with Rose Bengal antigen using Stirring
the titre of antibodies can be obtained by using specific Spectrum stick.
Brucella antigen suspensions. 6. Repeat these steps using the positive and negative controls
instead of serum sample.
Assay Principle 7. Gently rock the slide for 2 minutes (automatic rotator can
The smooth, killed stained Brucella antigen suspensions are also be used).
mixed with the patient’s serum. Specific antibodies to Brucella 8. Observe for agglutination after 2 minutes from beginning of
antigens if present in the patient serum will react with the antigen shaking.
suspension to produce an agglutination reaction. No agglutination
indicates the absence of specific antibodies to Brucella antigens. Interpretation of the results
Agglutination is a positive test result and indicates the presence
Reagent of specific antibodies to Brucella in the patient serum.
Spectrum Brucella Rose Bengal reagent contains ready to use
standardized, killed, stained, smooth specific antigen suspensions No agglutination is a negative test result and indicates absence
of Brucella having specific reactivity towards antibodies to of specific antibodies to Brucella in the patient serum.
Brucella Antigens.
Limitations
Reagent Storage and Stability 1. Agglutinins are not always produced in bacterial infections.
o
1. Store the reagents at 2 – 8 C (Do not freeze). 2. Cross reactions may occur in certain pathologies. For
2. The shelf life of reagent is as per the expiry date mentioned instance,Tularemia infections may produce agglutinins to
on the reagent vial labels. Each batch of reagents undergoes Brucella antigens.
rigorous quality control at various stages of manufacture for 3. Vaccinations may produce cross reacting antibodies.
its specificity, sensitivity and performance.
References
Note 1. J. G. Collee, J.P. Duguid, A G Fraser. Practical Medical
1. In-vitro diagnostic reagent for laboratory and professional Microbiology, 13 th Ed: 525-530.
use only. Not for medicinal use. 2. G.Galton, L.M.Jones, R.D.Angus, J.M.Verger. techniques
2. The reagent contains 0.01% thimerosal as preservative. for the Brucellosis laboratory,© INRA, Paris, 1988.
Avoid contact with skin and mucosa. On disposal 3. Felix A., (1942), Brit. Med. J., 11, 597.
flush with plenty of water. 4. Rose, J.E., Roepke, M.H., Am. J. Vet. Res. 18, 550-555
(1957).
Specimen Collection and Storage
1. No special preparation of the patient is required prior to
sample collection by approved techniques. Do not
use haemolysed samples.
2. Clean and dry glassware free from detergents must be used ORDERING INFORMATION
for sample collection. CATALOG NO. QUANTITY
3. Don’t heat inactive the serum.
4. Freshly collected serum is preferable, store samples at 2 – 711 001 50 test
8oC for 24 hours or frozen for several days. 711 002 100 test
711 003 100 test + Positive Control
Materials provided with the kit
1. Rose Bengal Brucella Antigen Suspension.
2. Positive Control (REF: 711 003).
242 243
TOXO LATEX KIT SYMBOLS IN PRODUCT LABELLING
A rapid latex agglutination test for qualitative EC REP Authorised Representative Temperature Limitation
and semi- quantitative detection of Toxoplasma IVD For in-vitro diagnostic use Use by/Expiration Date
gondii antibodies in serum LOT Batch Code/Lot number CAUTION. Consult instructions
REF: 729 001 100 test Consult instructions for use Manufactured by
REF: 729 002 200 test
Intended Use B) Semi-Quantitative Test

for qualitative and semi-quantitative detection of Toxoplasma gondii 1. Make serial two fold dilutions of the sample in normal saline
antibodies in serum solution.
2. Proceed for each dilution as in the qualitative method.
Background
Toxoplasma gondii is a specific protozoa in the genus Toxoplasma. Results and Interpretation
Toxoplasmosis, the disease of which T.gondii is the causative Negative result:
agent, is usually minor and self-limiting. the disease may also have No agglutination of the latex particles suspension within 4-6
serious effects on a fetus whose mother first contracts the disease minutes.
during pregnancy.
Positive result:
Principle An agglutination of the latex particles suspension will occur within
Toxo latex consists of an aqueous suspension of polystyrene 4-6 minutes, indicating an antibody concentration equal or more
particles coated with soluble purified antigens from Toxoplasma than 4 IU/mL.
gondii. If specific antibodies are present in the sample a clear The titer, in the Semi-quantitative method, is defined as the
visible agglutination will appear highest dilution showing a positive result.
MATERIALS PROVIDED Toxoplasma Ab Concentration

Spectrum TOXO latex kit contains the following components : Approximate anti-Toxoplasma concentration in the patient sample
is
• Toxo Latex Reagent : calculated as follows: 4xanti-Toxo Titer= IU/mL
Latex particles coated with soluble T.gondii antigen, pH,7.5
sodium azide 0.95 g/dL. Sensitivity
• Toxo Positive Control. The Sensitivity of the Kit is 4 IU/mL (3-6 IU/mL) under the
• Toxo Negative Control. recomended asay condition.
• Test slide.
• 20 G dispensing needle (20 ml/drop) Reference Value
Up to The 4 IU/mL
Storage & Stability
The reagents are stable up to the expiration date specified when Reference
o
stored at 2 – 8 C. 1. Young DS Effects of drugs on clinical laboratory test 4th ed.
AACC Press,1995.
Precautions and Warnings 2. Jacobs L.ADV Parasitol 1973;11;631-669
All human blood components used to prepare controls have been 3. Ruoss CF at al .The Journal of Obsterics and Gynecology of
tested for Hepatitis B surface antigen (HBsAg) and HTLV-III an- the British Commonwealth 1972 ;79:1115-1118
tibodies by FDA approved procedure and found to be non-reactive.
No known test method for HBsAg or HTLV-III antibodies offers total
assurance that a human derived product will not transmit hepatitis ORDERING INFORMATION
or HTLV-III virus. The user is therefore cautioned to handle reagents
CATALOG NO QUANTITY
as if being capable of transmitting these diseases.
729 001 100 test
Specimen Collection and Preservation 729 002 200 test
Use only fresh serum specimens , plasma samples are not suitable
o
for the test. Serum samples can be stored for 24 hrs at 2 – 8 C, for
o
longer Storage it is recommended to store the samples at -20 C
Haemolysis should be avoided.
Procedures
A) Qualitative
1. Bring reagents to room temperature.
2. Dispense 40ml of sample onto a single circle on the test slide.
3. Repeat step 2 for the positive and negative controls.
4. Spread the sample of each test specimen over the entire test
circle.
5. Shake the Toxo Latex reagent well.
6. With the needle suck up reagent sufficient to the testing
requirements.
7. Dispense one free-fall drop of the Latex reagent on each test
circle containing specimen.
8. Mix well and rotate slide slowly.
9. After 4-6 minutes check for agglutination .
244 245
F - H e m a t o l o g y,
Haemostasis Reagents
246 247
PT Reagent SYMBOLS IN PRODUCT LABELLING ISI
Or as international Normalized Ratio (INR). INR=(R) , where ISI = 5. Clotting time of patients on anticoagulant therapy depends
(SP-NORMOPLASTIN) EC REP Authorised Representative Temperature Limitation International Sensitivity index of the reagent (Refer to reagent vial upon the type and dosage of anticoagulant and also the time
IVD For in-vitro diagnostic use Use by/Expiration Date label) lag between the specimen collected and the last dose.
REF: 614 000 30 test 6. Turbid, icteric, lipemic or grossly hemolysed samples may
REF: 614 001 120 test *It is recommended by WHO that MNPT (mean normal PT) should be generate erroneous PT results.
REF: 614 002 240 test established for each lot of PT reagents by each laboratory, since PT 7. Glasswares and cuvettes used in the test must be scrupulously
REF: 614 003 80 test results are dependent on the combination of reagent lot, instrument clean and free from even traces of acids / alkalies or detergents.
o
and technique followed at each laboratory. Usually plasma from at 8. Plasma samples held at 4 – 8 C may undergo cold activation
least 20 normal healthy individuals should be used to establish the leading to a marked shortening of the PT.
Intended Use MNPT. The avarage of such PT results in seconds=MNPT. 9. The PT may be shortened during acute inflammatory conditions
Spectrum Diagnostics SP-NORMOPLASTIN reagent is intended for Transfer the blood into anticoagulated tubes, after detaching which are accompanied by increase in fibrinogen levels and also
prothrombin time (PT) determination. the needle from the syringe. Do not delay mixing blood with Expected Values by agent such as antihistamines, butabarbital, phenobarbital,
anticoagulant. Avoid foam formation during mixing.Mix exactly nine Normal values using SP-NORMOPLASTIN are between 10-14 caffeine, oral contraceptives and vitamin K. the PT may be
Background parts of freshly collected blood with one part of trisodium citrate (0.11 seconds. Between manual and turbo densitometric instrument prolonged by corticosteroides, EDTA, asparaginase, clofibrate,
The arrest of bleeding depends upon primary platelet plug formed mol/L, 3.2%). For occasional patients with hematocrit less than 20% results a variation of 1-2 seconds may be expected. For photo ethanol, tetracycline, aspirin and anticoagulants such as
along with the formation of a stable fibrin clot. Formation of this clot or greater than 55%, this ratio must be readjusted to ensure valid optical instruments, it is recommended that each laboratory must heparin and warfarin.
involves the sequential interaction of series of plasma proteins in result. Centrifuge immediately for 15 minutes at 1500 – 3000 rpm establish its own normal range. It is mandatory that each laboratory 10. It is important that each laboratory expresses the results in
a highly ordered and complex manner and also the interaction of (approximately 1500g) on laboratory centrifuge and transfer the must establish its own MNPT for each lot of SP-NORMOPLASTIN. terms of INR for patients on oral anticoagulant therapy for the
these complexes with blood platelets and materials released from plasma into a clean test tube. It should be ensured that the plasma is clinician to adjust the dosage based on INR.
the tissues. Tissue thromboplastin, in the presence of calcium, is free from platelets (PPP). Cap the test tubes to prevent deterioration Oral anticoagulant therapeutic range : INR = 2.0 – 3.5 11. Since the test uses platelet poor plasma, each laboratory
an activator which initiates the extrinsic pathway of coagulation, of samples. Plasma must be tested peferably immediately. However The use of INR’s enables direct comparison to be made between must calibrate the necessary force and time required during
o
which includes plasma coagulation factors VII, X, V, prothrombin and if the specimens are held at 2 - 4 C then they may be tested within all results on patient plasmas regardless of interlab variations or centrifugation to yield the PPP. Contamination of plasma with
fibrinogen. During oral anticoagulant therapy most of these factors 3 hours. reagent in question. excess platelets could falsely elevate levels of some of the
are depressed, as also during the deficiencies of clotting factor factors.
ISI
activity which may be hereditary or acquired. Prothrombin time Procedure The INR is calculated as INR = (R) 12. Homogenisation of SP-NORMOPLASTIN reagent suspension
determination is the preferred method for presurgical screening, Where ISI = Lot specific ISI for the reagent before use is important to achieve accurate and consistent
determination of congential deficiency of factors II, VII, X, V and for Manual Method results.
monitoring of patients on oral anticoagulant therapy and as a liver 1. Aspirate from the reagent vial enough reagent for immediate Patient PT
function test. testing requirements in a thoroughly clean and dry test tube And, R = References
(Plastic test tubes are preferred). Mean normal PT 1. Biggs R. and R.G. McFarlane: Human Coagulation and its
Assay Principle 2. Bring this reagent to room temperature before prewarming at disorders, Blackwell Scientific Publications, Oxford,1962.
o
Tissue thromboplastin in the presence of calcium activates the 37 C for testing purpose. Mean normal PT = Mean of the normal range that is specifically 2. Quick A.J., Hemorrhagic disease and thrombosis, 2nd Ed.,
o
extrinsic pathway of human blood coagulantion mechanism. When 3. Recap the reagent vial and replace immediately to 2 – 8 C. determined by each user laboratory for each lot of thromboplastin Philadelphia, Lee and Fibiger , 1966.
SP-NORMOPLASTIN reagent is added to normal antigoagulated 4. To a 12x75 mm tube add 0.1 ml of plasma (ppp) and place the reagent with specific instrument and techniques routinely used for 3. CRC Handbook Series in clinical laboratory Science, section1 :
o
plasma, the clotting mechanism is initiated, forming a solid gel clot tube in a water bath for 3 to 5 minutes at 37 C. patient testing. Haematology, Volume, 1980.
within a specified period of time. The time required for clot formation 5. To the tube forcibly add 0.2 ml of SP-NORMOPLASTIN reagent 4. E.A. Loeliger, A.M.H.P. Van den besselaar and S.M. Lewis,
would be prolonged if there is a deficiency of factors / factor activity
o
(prewarmed at 37 C for at least 3 minutes) and simultaneously Example: Reliability and Clinical impact of Normalization of Prothrombin
in the extrinsic pathway of the coagulation mechanism. start a stop watch. Shake the tube gently to mix contents. Patient PT result = 21 seconds Times in Oral anticoagulant control – F.K. Schattauer verlag
6. Gently tilt the tube back and forth and stop the stopwatch MNPT = 13.5 seconds. Gmbh 1985.
Reagent as soon as the first fibrin strand is visible and the gel / clot ISI of reagent = 1.5 5. Hull R., Hirsh H., jay R., et al., Difference intensities of oral
SP-NORMOPLASTIN is a liquid ready to use Calcium Throm- formation begins. Record the time in seconds. anticoagulant therapy in the treatment of proximalvein
boplastin reagent,which is derived from rabbit brain. Each batch of 7. Repeat steps 4-6 for a duplicate test on the same samples. 21.0 thrombosis, N. Engl. J . Med., 1982, 307, 1676-81.
reagents undergoes vigorous quality control at various stages of 8. Find the avarage of the duplicate test values. This is the R= = 1.5 6. W.H.O. Expert Committee on Biological standardization, No.
manufacture for its sensitivity and performance. Prothrombin Time (PT). 13.5 687, 1983.
1.5
Reagent Storage and Stability If a coagulation instrument is being used to perform the tests, the INR = (1.5) = 1.8
o
Store the reagent at 2 – 8 C (Do not freeze). the shelf life of reagent instrument manfacturers instructions must be strictly adhered to. SP-NORMOPLASTIN
is mentioned on vial label. The uncontaminated reagent is stable Alternatively the INR value can be read off directly from the ISI 1.5
o
for: 1 year at 2 – 8 C, 1 week at Calculation of Results SP-NORMOPLASTIN INR conversion table.
o o
18 – 25 C, 2 days at 37 C.
Manual Method Remarks ORDERING INFORMATION
Specimen Collection and Preparation of PPP The result may be reported directly in terms of the mean of the 1. It is recommended that controls with known factor activity CATALOG NO QUANTITY
Though no special preparation of the patient is required prior to double determination of PT of the test plasma in seconds. Or as a should be run simultaneously with each test series to validate
614 000 30 test
sample collection by approved techniques, it is preferable that ratio R: test run. 614 001 120 test
patients are not heavily exercised before blood collection. Fasting or 2. Incorrect mixture of blood and tri-sodium citrate, insufficient 614 002 240 test
only light non-fatty meals prior to blood collection provide samples Mean of the patient plasma PT in seconds prewarming of plasma and reagent, contaminated reagents, 614 003 80 test
with a desirable lower opacity. Withdraw blood without undue venous R= glassware etc. are potential source of errors.
stasis or frothing into a plastic syringe fitted with a short needle. MNPT for the reagent* 3. Oxalated plasma may induce prolonged clotting times.
o
The venipuncture must be a “clean” one and, if there is any difficulty, 4. Since the PT test functions correctly only at 37 ± 0.5 C,
take a new syringe and needle and try another vein. tempreature of all equipments must be calibrated daily.
248 249
NORMAL AND ABNORMAL SYMBOLS IN PRODUCT LABELLING Remarks
PLASMA CONTROLS FOR EC REP Authorised Representative
1. When used appropriately, PLASMATROL controls are
subjected to the limitation of the assay system deployed.
COAGULATION ASSAYS LOT
REF
Catalogue Number
for use
2. If proper values are not obtained it may indicate problems with
one or more variables of the assay system.
3. Stability of the reagent is dependent on storage and handling
REF: 618 001 ( HI ) 1 x 1 ml conditions. Since these can vary among laboratories, each
REF: 618 002 ( HII ) 1 x 1 ml laboratory should determine the stability of the reagent under
REF: 618 003 ( HI ) 2 x 1 ml usual operating conditions.
REF: 618 004 ( HI+HII ) 2 x 1 ml 4. Incorrect mixing of control plasma and reagent, insufficient
preparation of plasma/reagent, contaminated reagent, and
glassware etc. are a potential source of error.
5. Due to inter laboratory variations in techniques, standardization
Summary reparation Of The Reagent of test procedures and calibration of equipments, some
Spectrum Diagnostics PLASMATROL H-I and PLASMATROL 1. Reconstitute the control plasma with exactly 1 ml of bidistilled variation from assigned mean values may be expected.
H-II are two level human plasma controls that are suitable for use water, avoid using water containing preservatives.
as normal and abnormal control plasma for PT, APTT, TT and 2. Re-stopper the vial and allow to stand until, the hydration is
Fibrinogen testing using clot based methods. Coagulation controls complete (usually 5 - 7 minutes).
provide a means of day quality control in the laboratory for control of 3. Mix by gently swirling and inversion, avoiding froth formation.
accuracy and precision. Do not shake.
4. Allow to stand and equilibrate for a further 15 minutes before
Reagent use.
PLASMATROL is a stabilized and freeze dried preparation of 5. Use the reconstituted plasma within 3 hours of reconstitution.
selected human plasma with values determined and assigned for
specific clot based tests, which are lot specific. The plasma controls The Procedure
are assayed using spectrum Coagulation reagents. 1. Use the reconstituted PLASMATROL controls in the same
manner as freshly prepared citrated Platelet Poor Plasma from
Reagent Storage and Stability a patient.
o
Unopened vials should be stored at 2 – 8 C and are stable upon 2. Use the procedure as laid out in the ULTRAPLASTIN,
the expiry date mentioned on vial labels. After reconstitution the NORMOPLASTIN, and UNICELIN package inserts.
o
shelf life of the control plasma is 3 hours at 25 – 30 C, 8 hours when
o
stored at 2 – 8 C and one week at - 20 oC. For storage at - 20 C
o
Expected Values
avoid repeated freezing and thawing. 1. The expected value of specific assays are provided on the
assay value sheet accompanying each kit, and are lot specific.
Principle 2. The expected values are obtained using replicate assay of
The properties of the control plasma are similar to those of pooled each manufactured lot of PLASMATROL.
fresh plasmas. Since the plasma controls have assigned values 3. The individual laboratory values should fall within the expected
when substituted in place of a sample, in clot based coagulation values.
assays, they can be used for laboratory quality assurance. 4. It must however be noted that each laboratory should establish
its own normal values and reference range according to GLP.
Note
1. In-vitro diagnostic regent for laboratory and professional use
only.
NOT FOR MEDICINAL USE.
2. The source material used for preparation of the regent is
screened by third generation assays for HBsAg, HCV and HIV
antibodies and is found to be non-reactive. However handle
the material as if it is infectious, as no known test method can
assure that infectious agent are absent.
250 251
SP-Ultraplastin SYMBOLS IN PRODUCT LABELLING
Or as international Normalized Ratio (INR). INR=(R)ISI , where ISI
= International Sensitivity index of the reagent (Refer to reagent vial 7. Glasswares and cuvettes used in the test must be scrupulously
PT Reagent ISI 1.05 EC REP Authorised Representative
label) clean and free from even traces of acids / alkalies or detergents.
8. plasma samples held at 4 – 8oC may undergo cold activation
LOT Batch Code/Lot number CAUTION. Consult instructions *It is recommended by WHO that MNPT (mean normal PT) should be
REF: 622 001 ( 8 x 2 ml) 80 test established for each lot of PT reagents by each laboratory, since PT leading to a marked shortening of the PT.
REF: 622 002 ( 8 x 6 ml) 240 test results are dependent on the combination of reagent lot, instrument 9. The PT may be shortened during acute inflammatory conditions
and technique followed at each laboratory. Usually plasma from at which are accompanied by increase in fibrinogen levels and also
least 20 normal healthy individuals should be used to establish the
by agent such as antihistamines, butabarbital, phenobarbital,
MNPT. The avarage of such PT results in seconds=MNPT.
caffeine, oral contraceptives and vitamin K. the PT may be
Intended Use prolonged by corticosteroides, EDTA, asparaginase, clofibrate,
Expected Values
Spectrum Diagnostics SP-ULTRAPLASTIN is a sensitive Withdraw blood without undue venous stasis or frothing into a plastic Normal values using SP-ULTRAPLASTIN are between 11-15 ethanol, tetracycline, aspirin and anticoagulants such as
thromboplastin reagent intended for prothrombin time (PT) syringe fitted with a short needle. The venipuncture must be a “clean” seconds. Between manual and turbo densitometric instrument heparin and warfarin.
results a variation of 1-2 seconds may be expected. For photo
determination. one and, if there is any difficulty, take a new syringe and needle and optical instruments, it is recommended that each laboratory must 10. It is important that each laboratory expresses the results in
try anther vein. Trasfer the blood into anticoagulated tubes, after establesh its own normal range. It is mandatory that each laboratory terms of INR for patients on oral anticoagulant therapy for the
Background detaching the needle from the syringe. Do not delay mixing blood must establish its own MNPT for each lot of SP-ULTRAPLASTIN. clinician to adjust the dosage based on INR.
The arrest of bleeding depends upon primary platelet plug formed with anticoagulant. 11. Since the test uses platelet poor plasma, each laboratory
Oral anticoagulant therapeutic range : INR = 2.0 – 3.5
along with the formation of a stable fibrin clot. Formation of this clot The use of INR’s enables direct comparison to be made between must calibrate the necessary force and time required during
involves the sequential interaction of series of plasma proteins in Avoid foam formation during mixing.Mix exactly nine parts of all results on patient plasmas regardless of interlab variations or centrifugation to yield the PPP. Contamination of plasma with
a highly ordered and complex manner and also the interaction of freshly collected blood with one part of tri-sodium citrate (0.11 reagent in question. excess platelets could falsely elevate levels of some of the
these complexes with blood platelets and materials released from mol/l, 3.2%). For occasional patients with hematocrit less than The INR is calculated as INR = (R) ISI factors.
Where ISI = Lot specific ISI for the reagent
the tissues. 20% or greater than 55%, this ratio must be readjusted to ensure 12. Homogenisation of SP-ULTRAPLASTIN reagent suspension
Tissue thromboplastin, in the presence of calcium, is an activator valid result. Centrifuge immediately for 15 minutes at 1500 – 3000 before use is important to achieve accurate and consistent
which initiates the extrinsic pathway of coagulation, which includes rpm(approximately 1500g) on laboratory centrifuge and transfer the Patient PT results.
plasma coagulation factors VII, X, V, prothrombin and fibrinogen. plasma into a clean test tube. It should be ensured that the plasma is And, R =
During oral anticoagulant therapy most of these factors are free from platelets (PPP). Cap the test tubes to prevent deterioration Mean normal PT References
depressed, as also during the deficiencies of clotting factor activity of samples. Plasma must be tested peferably immediately. However 1. Biggs R. and R.G. McFarlane: Human Coagulation and its
Mean normal PT = Mean of the normal range that is specifically
which may be hereditary or acquired. if the specimens are held at 2 - 4 oC then they may be tested within disorders, Blackwell Scientific Publications, Oxford, 1962.
determined by each user laboratory for each lot of thromboplastin
Prothrombin time determination is the preferred method for 3 hours. reagent with specific instrument and techniques routinely used for 2. Hirsh J., Dalen J.E., Deykin D., poller L.: Oral Anticoagulants
presurgical screening, determination of congential deficiency of patient testing. : Mechanism of Action, Clinical Effectiveness and Optical
factors II, VII, X, V and for monitoring of patients on oral anticoagulant Procedure Therapeutic Range, Chest : 1995 : 108 (suppl.) : 231S-246S.
therapy and as a liver function test. Manual Method Example: 3. W.H.O. Expert Committee on Biological standardization, No.
Patient PT result = 21 seconds
1. Aspirate from the reagent vial enough reagent for immediate MNPT = 13.5 seconds. 687, 1983.
Assay Principle testing requirements in a thoroughly clean and dry test tube ISI of reagent = 1.1 4. Colman R., Hirsh J.: Haemostasis & Thrombosis, J.B. Lippincott
Tissue thromboplastin in the presence of calcium activates the (Plastic test tubes are preferred). Company, 3rd Ed., 1994.
extrinsic pathway of human blood coagulantion mechanism. When 2. Bring this reagent to room temperature before prewarming at 21.0
SP-ULTRAPLASTIN reagent is added to normal antigoagulated 37 oC for testing purpose. R = =1.5
plasma, the clotting mechanism is initiated, forming a solid gel clot 3. Recap the reagent vial and replace immediately to 2 – 8oC. 13.5
within a specified period of time. The time required for clot formation 4. To a 12x75 mm tube add 0.1 ml of plasma (ppp) and place the INR = (1.5)1.1 = 1.56 CATALOG NO QUANTITY
would be prolonged if there is a deficiency of factors / factor activity tube in a water bath for 3 to 5 minutes at 37 oC.
622 001 8 x 2 ml
in the extrinsic pathway of the coagulation mechanism. 5. To the tube forcibly add 0.2 ml of SP-ULTRAPLASTIN reagent Alternatively the INR value can be read off directly from the SP- 622 002 8 x 6 ml
(prewarmed at 37 oC for at least 3 minutes) and simultaneously ULTRAPLASTIN INR conversion table.
Reagent start a stop watch. Shake the tube gently to mix contents.
SP-ULTRAPLASTIN is a liquid ready to use Calcium Thromboplastin 6. Gently tilt the tube back and forth and stop the stopwatch Remarks
reagent,which is derived from rabbit brain. Each batch of reagents as soon as the first fibrin strand is visible and the gel / clot 1. it is recommended that controls with known factor activity
undergoes vigorous quality control at various stages of manufacture formation begins. Record the time in seconds. should be run simultaneosly with each test series to validate
for its sensitivity and performance. 7. Repeat steps 4-6 for a duplicate test on the same samples. test run.
8. Find the avarage of the duplicate test values. This is the 2. incorrect mixture of blood and tri-sodium citrate, insufficient
Reagent Storage and Stability Prothrombin Time (PT). prewarming of plasma and reagent, contaminated reagents,
Store the reagent at 2 – 8oC (Do not freeze) . the shelf life of reagent glassware etc. are potential source of errors.
is mentioned on vial label. The uncontaminated reagent is stable for If a coagulation instrument is being used to perform the tests, 3. Oxalated plasma may induce prolonged clotting times.
the instrument manfacturers instructions must be strictly ad-
: 1 year at 2 – 8oC, 1 week at 18 – 25oC, 2 days at 37oC. 4. Since the PT test functions correctly only at 37 ± 0.5 oC,
hered to.
tempreature of all equipment must be calibrated daily.
Specimen Collection and Preparation of PPP Calculation of Results 5. clotting time of patients on anticoagulant therapy depends upon
Though no special preparation of the patient is required prior to Manual Method the type and dosage of anticoagulant and also the time lag
sample collection by approved techniques, it is preferable that The result may be reported directly in terms of the mean of the between the specimen collected and the last dose.
double determination of PT of the test plasma in seconds.
patients are not heavily exercised before blood collection. Fasting or Or as a ratio R: 6. turbid, icteric, lipemic or grossly hemolysed samples may
only light non-fatty meals prior to blood collection provide samples generate erroneous PT results.
with a desirable lower opacity. Mean of the patient plasma PT in seconds
R=
MNPT for the reagent*
252 253
APTT Reagent SYMBOLS IN PRODUCT LABELLING Calculation and reporting of results
EC REP Authorised Representative Temperature Limitation a) The result may be reported directly in terms of the mean of the
(SP- UNICELIN) IVD
LOT
double determination of the APTT of the test plasma

OR
REF: 626 001 150 test
REF: 626 002 160 test b) as a ratio R as follows:
REF: 626 003 240 test
APTT of patient plasma (in seconds)
R=
APTT of FNP (in seconds)
Intended Use Specimen Collection and Preparation
Spectrum Diagnostics SP- UNICELIN reagent is intended for partial No special prepartion of the patient is required prior to sample Expected Values
Thromboplastin (APTT) determination using ellagic acid, as an collection. Withdraw blood without doing venous stasis and avoid Normal values are between 22-34 seconds.
activator. haemolysis. The veinpuncture must be a clean one and, if there is
any difficulty, take a new syringe and needle and try another vein. Remarks
Background Mix exactly nine parts of freshly collected blood with one part of 1. Each laboratory must establish its own normal population
The arrest of bleeding depends upon primary platelet plug formed trisodium citrate (0.11mol/L, 3.2 %). Centrifuge immediately for 15 range as well as normal and abnormal range.
along with the formation of a stable fibrin clot. Formation of this clot minutes at 3000 rpm and transfer the plasma into a clean test tube. 2. Clotting time of patients on anticoagulant therapy depends
involves the sequential interaction of a series of plasma proteins in Plasma must be tested within 3 hours of blood collection. upon the type and dosage of anticoagulant and also the time
a highly ordered and complex manner and also in the interaction of lag between the specimen collection and the dose.
these complexes with blood platelets and materials released from Pooled Plasma 3. Abnormalities of coagulation factor VII, factor XIII, and platelets
the tissues. Prepare a fresh normal plasma pool (FNP) from at least five normal are not detected by this method.
Activated Partial Thromboplastin Time (APTT) is prolonged with healthy donors and process as above. Plasma must be tested within 4. Platelet factor IV, a heparine-nutralising factor can be released
a deficiency of coagulation factors of the intrinsic pathway of the 3 hours of blood collection. due to platelet aggregation or damage. In order to prevent this
human coagulation mechanism such as factor XII, XI, IX, VIII, X, phenomenon in-vitro the specimen should be collected with a
V, II, and Fibrinogen. Determination of APTT helps in estimating Additional regent minimum of trauma.
abnormality in most of the clotting factors of the intrinsic pathway 0.025 mol/L calcium chloride (available from Spectrum Diagnostics 5. Decrease in APTT time is observed in males under estrogen
and is also a sensitive procedure for generating heparin response upon request). therapy and oral contraceptive administration in females.
curves for monitoring heparin therapy.
Procedure References
Assay Principle 1. Before use, the reagent should be mixed well by gentle swirling 1. Biggs, R.ed.: Human Blood Coagulation, Haemostasis and
Cephaloplastin activates the coagulation factors of the intrinsic , do not shake. Thrombosis, Blackwell Scientific Publications, Oxford, England,
pathway of the coagulation mechanism in the presence of 2. Aspirate from the reagent vial enough reagent for the immediate 1972.
calciumions. APTT is prolonged by deficiency of one or more of test requirement in extremely clean and dry test tube. Bring 2. Hoffmann, J.J.M.L and Neulendijk P.N.: Thrombos.Haemosta.
these clotting factors of the intrinsic pathway and in the presence of this reagent to room temperature before prewarming at 37 °C (Stuttgard) 39, 640 (1978).
coagulation inhibitors like heparin. for testing procedure. The calcium chloride solution should be
brought to 37 °C before use.
Reagent 3. To 12 x 75 mm test tube, add 0.1 ml test plasma and 0.1 ml SP- ORDERING INFORMATION
SP-UNICELIN is a liquid ready-to-use activated cephaloplastin UNICELIN. Shake tube briefly to mix the reagent and plasma, CATALOG NO QUANTITY
reagent for the determination of APTT. It is phospholipids preparation place tube at 37 °C for 3 minutes.
626 001 150 test
derived from rabbit brain with ellagic acid as an activator. 4. Add forcibly o.1 ml prewarmed calcium chloride and 626 002 160 test
simultaneously start stop watch. Shake tube briefly to mix 626 003 240 test
Reagent Storage and Stability contents, keep at 37°C for 20 seconds.
Store the reagent at 2 – 8 °C. Never freez the reagent. The reagent 5. Following 20 seconds incubation, remove the tube, gently tilt
is stable for 1 year at 2–8 °C, 1 week at 18–25 °C, 2 days at 37 °C. back and forth until a gel clot forms, stop the watch and ,record
the time.
Note 6. Repeat for a duplicate test using the same test plasma.
1. Avoid exposure of the reagent to elevated temperature and 7. Find the average from the duplicate test values. This is the
contamination. Activated Partial Thromboplastin Time (APTT of patient plasma)
2. Immediately replace cap after use and store at recommended 8. Similarly repeat the steps 2-4 twice , and record values using
temperature. FNP in place of test plasma ( APTT of FNP).
3. Reagent contain 0.01 g/dL Thimerosal as a preservative. Avoid
contact with skin and mucosa. On disposal, flush with plenty
of water
254 255
G-blood group
256 257
Spectrum Anti-A, SYMBOLS IN PRODUCT LABELLING Slide and Tube tests
Anti-B,Anti-AB EC REP Authorised Representative

Agglutination is a positive test result and indicates the presence of A CATALOG NO QUANTITY
and/or B antigen. Do not interpret peripheral drying or fibrin strands
Monoclonal (IgM) LOT
REF
Catalogue Number
for use
as agglutination. No agglutination is a negative test results and
Anti A
Blood Grouping Reagents Consult instructions for use Manufactured by

indicates the absence of A and/or B antigen. 810 001
810 002
1 x 10 ml
10 x 10 ml
Anti-A Anti-B Anti-AB

Remarks Anti B
1. (a) Spectrum Anti-A, Anti-B and Anti-AB reagent do not show
810 001 1 x 10 ml 814 001 1 x 10 ml 816 001 1 x 10 ml 814 001 1 x 10 ml
810 002 10 x 10 ml 814 002 10 x 10 ml 816 002 10 x 10 ml a reaction with crypt antigens (T, Tn,Tk activated cells). (b)
814 002 10 x 10 ml
Spectrum Anti-B is truly negative reacting with acquired B
characteristics. Anti AB
2. In the tube test procedure, it is recommended that tubes with
816 001 1 x 10 ml
Intended Use Reagent Storage and Stability negative reactions should be recentrifuged and results read
o
816 002 10 x 10 ml
Spectrum Diagnostics Anti-A , Anti-B and Anti-AB reagent is intended 1. Store the reagent at 2-8 C. Don’t freeze. again after 5 minutes so that weak antigens are not overlooked.
for the detection of Blood groups A, B and AB in Human Blood. 2. The shelf life of reagent is as per the expiry date mentioned on 3. As undercentrifugation or overcentrifugation could lead to
the reagent vial label erroneous results, it is recommended that each laboratory
Background calibrate its own equipment and determine the time required for
Monoclonal antibodies are derived from hybridoma cell lines, created Specimen Collection and Storage achieving the desired results.
by fusing mouse antibody producing B lymphocytes with mouse No special preparation of the patient is required prior to sample 4. Results of forward grouping obtained by using Anti-A, Anti-B,
myeloma cells. Each hybridoma cell line produces homogenous collection by approved techniques. Samples should be stored at Anti- AB reagents should always be reconfirmed by performing
o
antibodies of only one immunoglobulin class, which are identical in 2-8 C if not tested immediately. Do not use haemolysed samples. reverse grouping with known red cells.
their chemical structure and immunological activity. Anticoagulated blood using various anticoagulants should be tested 5. It is strongly recommended that red cells with known ABO
Human red Blood cell antigens can be divided into four groups within the below mentioned time period: characteristics should be occasionally run, preferably on a daily
A, B, AB and O depending on the presence or absence of the EDTA or heparin : 2 days basis so as to control reagent performance and validate test
corresponding antigens on the red blood cells. Approximately 41% Sodium citrate or Sodium oxalate : 14 days results.
of the Caucasian population have the A antigen, 9% have the B ACD or CPD : 28 days 6. After usage the reagents should be immediately recapped and
o
antigen, 4% have both A and B antigens, while remaining have Clotted whole blood should be tested within 14 days. replaced to 2-8 C storage.
neither A nor B antigen. 7. The label minimum titre claim is based on A1 group cells
Additional Material Required For Slide And Tube Tests for Spectrum Anti-A reagent, B group cells for Spectrum
Assay Principle Glass slides (50x75 mm), Test tubes (12x75 mm), Pasteur Anti-B reagentand A1B cells for Spectrum Anti AB reagent.
Human red blood cells possessing A and/or B antigen will agglutinate pipettes,isotonic saline, Centrifuge, timer, mixing sticks. This is based on titration procedure as recommended by the
in the presence of antibody directed towards the antigen. manufacturer, Any deviation in test procedure could result in
Agglutination of red blood cells with Anti-A, Anti-B, Anti-A,B Procedure variable results.
reagents is a positive test result and indicates the presence of the Bring reagent and samples to room temperature before testing.
corresponding antigen. Warranty
Absence of agglutination of red blood cells with Anti-A, Anti-B, Anti- Slide Test This product is designed to perform as described on the label and
A,B reagents is a negative test result and indicates the absence of 1. Place one drop of Spectrum Anti-A or Anti-B or Anti-AB reagent package insert. The manufacturer disclaims any implied warranty of
the corresponding antigen. on a clean glass slide. use and sale for any other purpose.
2. To each reagent drop, add one small drop (50 ml) of whole
Note blood. References
1. In-vitro diagnostic reagent for laboratory and professional use 3. Mix well with a mixing stick uniformly over an area of 1. Kohler C. & Milstein C. (1975), Continuous cultures of fused
only. Not for medicinal use. approximately 2.5 cm². Rock the slide gently, back and forth. cells secreting antibody of predefined specificity, Nature, 256,
2. The reagent contains sodium azide 0.1% as preservative. 4. Observe for agglutination macroscopically at two minutes. 495-497.
Avoid contact with skin and mucosa. On disposal flush with 2. Lee H.H., Rouger P, Germain C., Muller A. & Salmon C.
large quantities of water. Tube Test (1983),The production and standardisation of monoclonal
3. Extreme turbidity may indicate microbial contamination or 1. Prepare a 2-3% suspension of the red cells to be tested in antibodies as AB blood group typing reagents. Symposium
denaturation of protein due to thermal damage. Such reagent Isotonic saline. of International Association of Biological Standardisation on
should be discarded. 2. Place one drop of Spectrum Anti-A, Anti-B, Anti-AB into Monoclonal antibodies.
4. 4Spectrum blood grouping reagents are not from human correspondingly labeled test tubes. 3. Human Blood Groups by Geoff Daniels, 1st Ed., Blackwell
sources,hence contamination due to HBsAg and HIV is 3. Pipette into each of the test tubes, one drop ( 50ml) of the test Science,Oxford 1995.
practically excluded. red cell suspension and mix well. 4. HMSO, Guidelines for the Blood Transfusion Services, 2nd Ed.,
4. Centrifuge for 1 minute at 1000 rpm (125 g) or 20 seconds at 1994.
Reagents 3400 rpm (1000 g) or incubate at room temperature for 20-30
Spectrum Anti-A, Anti-B and Anti-AB are ready-to-use reagents minutes.
prepared from supernatants of mouse hybridoma cell cultures. 5. Gently resuspend the cell button, observing for agglutination
These antibodies of immunglobulin class lgM are a mixture of macroscopically.
several monoclonal antibodies of the same specificity but having the
capability of recognising different epitopes of the human red blood Inerpretation Of Results
cell antigens A and B. Each batch of reagent undergoes quality
control at various stages of manufacture for its specificity, avidity
and performance.
258 259
Anti-D (Rho) Plus SYMBOLS IN PRODUCT LABELLING u
D Test Procedure Bibliography
Monoclonal IgM & IgG EC REP Authorised Representative
1. Prepare a 5% suspension of the red cells to be tested in 1. Kohler C. & Milstein C. (1975), Continuous cultures of fused
isotonic saline. cells secreting antibody of predefined specificity, Nature, 256,
Blood Grouping Reagent LOT
REF
Catalogue Number
for use
2. Place one drop of Spectrum Anti-D (Rho) plus into labelled 495-497.
test tube. 2. Lee H.H., Rouger P, Germain C., Muller A. & Salmon C.
REF: 822 001 1 x 10 ml 3. Add to the test tube, one drop of the 5 % red cell suspension (1983),The production and standardisation of monoclonal
REF: 822 002 10 x 10 ml and mix well, incubate at 37 ºC for 15 minutes antibodies as AB blood group typing reagents. Symposium
4. Wash the contents of the tube at least three times, with isotonic of International Association of Biological
saline and decant completely after the last wash. Standardisation on Monoclonal antibodies.
Intended Use 5. Add 2 drops of Anti Human Globulin reagent and mix well. 3. Human Blood Groups by Geoff Daniels, 1st Ed., Blackwell
Spectrum Diagnostics Anti-D (Rho) plus reagent is intended for the plus reagent is a blend of IgM and IgG class of Anti-D (Rho) 6. Centrifuge for one minute at 1000 rpm (125 g) or 20 seconds at Science, Oxford 1995.
detection of Rho (D) Type in human Blood. monoclonals, a characteristic which accords versatility to the reagent. 3400 rpm ( 1000 g). 4. HMSO, Guidelines for the Blood Transfusion Services, 2nd Ed.,
It gives an avid saline reacting slide / tube test reagent the capability 7. Very gently, resuspend the cell button and observe for 1994.
Background of detecting Du (weak / partial D´s) in the Anti-human globulin phase. agglutination macroscopically. 5. AABB Technical manual, 13th Ed., 1999.
Monoclonal antibodies are derived from hybridoma cell lines, created Each batch of reagent undergoes quality control at various stages of
by fusing mouse antibody producing B lymphocytes with mouse manufacture for its specificity, avidity and performance. Inerpretation Of Results ORDERING INFORMATION
myeloma cells or are derived from a human B cell line through EBV CATALOG NO QUANTITY
transformation. Each hybridoma cell line produces homogenous Reagent Storage and Stability Slide and Tube tests
o 822 001 1 x 10 ml
antibodies of only one immunoglobulin class, which are identical 1. Store the reagent at 2-8 C. Don’t freeze. 822 002 10 x 10 ml
in their chemical structure and immunological activity. Human red 2. The shelf life of reagent is as per the expiry date mentioned on (a) Agglutination is a positive test result and indicates the presence
Blood cells are classified as Rho (D) positive or Rho (D) negative the reagent vial label of D (Rho) antigen. Do not interpret peripheral drying or fibrin
depending on the presence or absence of Rho (D) antigen on them. Specimen Collection and Storage strands as agglutination. No agglutination is a negative test result
Approximately 85 % of the Caucasian population is Rho (D) positive. No special preparation of the patient is required prior to sample and indicates absence of D (Rho) antigen.
The Du phenotype is a traditional definition to describe the weak collection by approved techniques. Samples should be stored at (b) Cord cells heavily sensitised with Anti- D (Rho) may give
/ partial D´s that can be detected with spectrum anti-D Rho (IgM 2-8 oC if not tested immediately. Do not use haemolysed samples. a false negative immediate spin test rresult
& IgG). About 60 % of the Dus (weak / partial D´s) may react with Anticoagulated blood using various anticoagulants should be tested
Spectrum Anti-D (Rho) plus reagent in slide test and about 90 % within the below mentioned time period: Procedure
may be detected by the tube technique. EDTA or HEPARIN : 2 days (a) Agglutination with the Anti Human Globulin reagent and no
Sodium citrate or Sodium oxalate : 14 days agglutination with the control test indicates the presence of the
Assay Principle ACD or CPD : 28 days Du antigen (weak / partial D´s). No agglutination with both indicates
Human red blood cells possessing D antigen will agglutinate in the Clotted whole blood should be tested within 14 days. the absence of the Du antigen (weak/partial D´s)
presence of antibody directed towards the antigen. Agglutination of (b) Red cells demonstrating a positive direct antiglobulin test cannot
red blood cells with Spectrum Anti-D (Rho) plus reagent is a positive Additional Material Required For Slide And Tube Tests be accurately tested for Du antigen (weak/partialD´s)
test result and indicates the presence of D (Rho) antigen. No Glass slides (50x75 mm), Test tubes (12x75 mm), Pasteur
agglutination with Spectrum Anti-D (Rho) plus reagent is a negative pipettes,isotonic saline, Centrifuge, timer, mixing sticks. Anti Human Remarks
test result and indicates the absence of the D (Rho) antigen. All Globulin (Coombs) reagent 1. As undercentrifugation or overcentrifugation could lead to
negative test results should be further tested for Du(weak/partialD´s) erroneous results, it is recommended that each laboratory
by performing the Du test procedure, as described later. Procedure calibrate its own equipment and determine the time required for
Bring reagent and samples to room temperature before testing. achieving the desired results.
Note 2. It is strongly recommended that red cells with known Rho (D)
1. In-vitro diagnostic reagent for laboratory and professional use Slide Test positive and Rho (D) negative be occasionally run, preferably
only. Not for medicinal use. 1. Place one drop of Spectrum Anti-D (Rho) plus reagent on a on a daily basis so as to control reagent performance and
2. The reagent contains sodium azide 0.1% as preservative. clean glass slide. validate test results.
Avoid contact with skin and mucosa. On disposal flush with 2. Add one equal drop of whole blood on the slide. 3. After usage the reagents should be immediately recapped and
large quantities of water. 3. Mix well with a mixing stick uniformly over an area of replaced to 2-8 °C storage.
3. Extreme turbidity may indicate microbial contamination or approximately 2.5 cm². Rock the slide gently, back and forth.
denaturation of protein due to thermal damage. Such reagent 4. Observe for agglutination macroscopically at two minutes. Warranty
should be discarded. This product is designed to perform as described on the label and
4. Spectrum Anti-D (Rho) plus reagent is not from human Tube Test package insert. The manufacturer disclaims any implied warranty of
source,hence contamination due to HBs Ag and HIV is 1. Prepare a 5% suspension of the red cells to be tested in use and sale for any other purpose.
practically excluded. isotonic saline.
2. Place one drop of Spectrum Anti-D (Rho) plus into the labelled
Reagents test tubes.
Spectrum Anti-D (Rho) plus is ready to use reagent prepared from 3. Pipette into each of the test tubes, one drop of the 5% red
supernatants of cell cultures with antibody producing B lymphocytes cell suspension and mix well,
obtained through EBV transformation and is a blend of monoclonal 4. Centrifuge for 1 minute at 1000 rpm (125 g) or 20 seconds at
antibodies of immunoglobulin class IgM and IgG. These antibodies 3400 rpm (1000 g).
are a mixture of several monoclonal antibodies of the same 5. Gently resuspend the cell button, observing for agglutination
specificity but having the capability of recognising different epitopes macroscopically
of the human red blood cell antigen D (Rho). Spectrum Anti-D (Rho)
260 261
ANTI-HUMAN GLOBULIN SYMBOLS IN PRODUCT LABELLING Indirect Test - Tube method
EC REP Authorised Representative Temperature Limitation 1. Wash the red cells 4 times in large volumes in physiological
SERUM IVD
LOT
saline. Decant completely the last wash
2. Re-suspend the cells to 5% suspension in physiological saline
(COOMBS) REF Catalogue Number
Consult instructions for use
for use
Manufactured by
3. Place in a small Test tube:
2 volumes of Spectrum anti-human globulin reagent
REF: 819 001 (1 X 10 ml) 1 volume of 3% suspension test red cells
REF: 819 002 (10 X 10 ml) 4. Mix well and centrifuge at 1000 rpm (100 RDF) for 1 minute
5. Agitate the tube gently and examine macroscopically for
agglutination. Negatives can be checked microscopically
Intended Use Indirect Test - Tube method Notes

Spectrum Diagnostics Anti-Human Globulin Serum is intended for 1. Prepare 2 -4 % suspension of the cells to be used in the test in 1. Appropriate positive and negative controls must be used with
the in-vitro detection of antibody coating on human physiological saline (0.85% Nacl pH 7.0) each test or batch of test
erythrocytes 2. Place in small Test tube: 2. Spectrum anti-human globulin reagent is suitable for use with
Two volumes of serum to be tested automated Coombs washing equipment
Background 1 volume of 3% red cell suspension 3. This reagent is prepared by blending the serum from rabbits
Antibodies immunoglobulins may become attached to human red 1 volume of 22% or 30 % spectrum Bovine albumin which have been immunized with different human globulin
o
cells either “ in-vivo” or “in-vitro” 3. Mix well and incubate at 37 C for 30 minutes preparations
“In-vivo” coating can occur if the body produces an auto-antibody 4. Wash the cells 4 times in large volumes of physiological saline 4. Preservative :0.1% sodium azide
o
against a self antigen located on its own red cells. Decant completely the last wash store at 2 - 8 C
“In-vitro” coating can occur during blood grouping tests compatibility 5. Add 2 volumes of Spectrum anti-human globulin reagent
testing prior to transfusion or when testing to detect and investigate 6. Mix well and centrifuge at 1000 rpm (100 RDF) for 1 minute
atypical antibodies 7. Agitate the tube gently and examine macroscopically for
agglutination. Negatives can be checked microscopically
Recommended Procedure
Direct Test - Tile method ORDERING INFORMATION
Indirect Test - Tile method 1. Wash the red cells to be tested in large volumes of physiological CATALOG NO QUANTITY
1. Prepare 2 - 4 % suspension of red cells to be used in the test in saline. Decant completely the last wash
819 001 1 x 10 ml
physiological saline (85% Nacl pH 7.0) 2. Prepare a 3% suspension of washed red cells in physiological 819 002 10 x 10 ml
2. Place in a small Test tube: saline
Two volumes of serum to be tested 3. Mix on a clean tile or slide:
1 volume of 3% red cell suspension 1 drop of Spectrum anti-human globulin reagent
1 volume of 22% or 30 % spectrum Bovine albumin 1 drop of 3 % red cell suspension
o
3. Mix well and incubate 37 C for 30 minutes 4. Allow to stand at room tempereature for 5 minutes
4. Wash the cells 4 times in large volumes in physiological saline 5. Rock the tile gently and examine for agglutination over a light
Decant completely the last wash source
5. Re-suspend the cell to a 3% suspension in physiological saline
6. Mix on a clean tile or slide: 1 volume of Spectrum anti-human
globulin reagent 1 volume of 3 % suspension washed cells
7. Allow to stand at room tempereature for 5 minutes
8. Rock the tile gently and examine for agglutination over a light
source
262 263
BOVINE SERUM ALBUMIN SYMBOLS IN PRODUCT LABELLING Technique - Antibody Titration Procedure
EC REP Authorised Representative Temperature Limitation 1. Prepare doubling dilution of test serum in either normal group ORDERING INFORMATION
(BSA) IVD
LOT
AB serum or 6% bovine albumin (the latte can be prepared CATALOG NO QUANTITY
by mixing 1 part spectrum 30% bovine albumin with 4 parts
isotonic buffered saline BSA 22%
BSA 22% BSA 30% 2. Prepare a 2% suspension of appropriate washed red cells in
922 001 1 x 10 ml
REF: 922 001 (1 X 10 ml) REF: 924 001 (1 X 10 ml) spectrum 22% or 30% bovine albumin 922 002 10 x 10 ml
REF: 922 002 (10 X 10 ml) REF: 924 002 (10 X 10 ml) Technique - Albumin Replacement Method 3. Add 1 volume of 2% cell suspension to 1 volume of each serum
1. Prepare a 2 - 3% suspension of red cells in isotonic buffered dilution mix well and incubate at 37 oC for 15 - 60 minutes BSA 30%
saline (pH 6.9) 4. Centrifuge at 900 - 1000 rcf for 30 seconds 924 001 1 x 10 ml
2. Place in a glass test tube 5. Gently resuspend the cell button and Examine for agglutination. 924 002 10 x 10 ml
Intended Use 1 Volume of serum or plasma Record result
Spectrum Diagnostics Bovine Serum Albumin is intended for 1 Volume of 2-3% cell suspension 6. An antiglobulin test may be preformed on those cells showing
o
the in-vitro use 3. Mix well and incubate at 37 C for 45 - 90 minutes. weak or negative results
4. With a fine pipette remove the supernatant saline-serum
Background mixture, leaving the button of red cells * Alternatively a time appropriate for the centrifuge being used may
Incomplete antibodies have the ability to combine with their specific 5. Add one volume of spectrum 22% Bovine Albumin.Taking care be determined i,e., that which produces the strongest reaction of
antigens in the first stage of agglutination but will not produce not to disturb the cell button antibody with antigen-positive cells, yet allows easy resuspension of
o
visible agglutination without the use of special techniques. Addition 6. without mixing reincubate at 37 C for 15-30 minutes antigen-negative cells
of bovine albumin to the cell suspension enable some of these 7. Examine for agglutination. Reactions may be examined with an
antibodies to complete the second stage of agglutination . Albumin optical aid, or microscopically. Record results. Stability of the reaction
has been shown to enhance the sensitivity of the indirect antiglobulin Following the recommended procedures all tests should be read
test for some antibody specificities specification. Technique - Albumin Displacement Method immediately and results interpreted without delay. Delays in reading
1. Follow steps 1-3 of albumin replacement method. or delays in completion of washing steps where appropriate may
Reagents 2. Upwardly displace the supernatant saline-serum mixture by result in dissociation of antigen antibody complexes, leading to false
Spectrum 22% and 30% Bovine albumin reagents are prepared carefully allowing one volume of Spectrum 30% Bovine albumin negative or weak positive reactions
from bovine serum albumin. The polymer content of spectrum to run down the inside wall of the test tube.
22% polymer Enhanced Bovine Albumin is increased naturally 3. Follow steps 6 and 7 of the Albumin Replacement Method Limitations
by a process modification . No artificial avidity enhancers or high Bovine albumin will not enhance the reactivity of all blood group
molecular weight agglutination potentiators are added to BSA. Technique - Albumin Mix Method antibodies.
These reagents do not contain sodium caprylate. Sodium azide at 1. Prepare a 2 - 3% suspension of red cells in isotonic buffered Bovine albumin solutions should not be used as negative controls
0.1 % is added as a preservative . these reagents should be used as saline (pH 6.9) for potentiated IgG blood grouping reagents. False positive or
supplied by the methods described ; their suitability for use in other 2. Place in a glass test tube false negative results may occur due to improper technique or
techniques must be determined by the user. 1 Volume of serum or plasma contaminated test materials.
1 Volume of 2-3% cell suspension
Precautions 2 Volumes of Spectrum 22% Bovin Albumin Specific Performance Characteristics
o o
For in vitro diagnostic use only . Store at 2-8 C when not in use . Do 3. Mix well and incubate at 37 C for 15 - 60 minutes. Spectrum 22% and 30% Bovine Albumin solution have been shown
not freeze or expose to elevated temperatures .market turbidity may 4. Centrifuge at 900 - 1000 rcf for 30 seconds. to enhance agglutination of Rh and other antibodies when used
indicate reagent deterioration or contamination. Spectrum Bovine 5. Gently resuspend the cell button and Examine for agglutination. according to insert methodologies. Each lot is tested to assure
Albumin solutions are derived from accredited and inspected herds Record result specificity in an antibody- free system with red cells Known to
from areas where the risk of bovine spongiform encephalopathy process the most frequently inherited blood group antigens.
is negligible. Additionally, during the manufacturing process the Technique - Indirect Antiglobulin Test
reagents are subjected to conditions of high temperature and low 1. Follow steps 1-3 of albumin mix method
PH for extended periods. Such procedures have been shown to 2. Wash the cells 3-4 times in isotonics buffered saline, decanting
completely inactive BSE-like agent the saline completely after each wash
3. Add two volumes of Spectrum Anti Human Globulin to the dry
Specimen Collection cell button
Fresh serum obtained from a fully clotted specimen should be used 4. Mix gently and centrifuge at 900- 100 rcf for 15 seconds
in compatibility or antibody identification procedures. Red cells 5. Gently resuspend the cell button and Examine for agglutination.
obtained from samples with or without anticoagulants can be used in Record result
antigen detection tests.Draw a blood specimen using an acceptable 6. Confirm validity of all negative reactions by using IgG sensitised
phlebotomy technique. Testing should be performed as soon as red cells, such as spectrum Coombs Control cells
o
possible. If testing is delayed store samples at 2 - 8 C. Serum or
Plasma can be separated from the cells and frozen
264 265
Low Ionic Strength (LISS) SYMBOLS IN PRODUCT LABELLING
EC REP Authorised Representative Temperature Limitation Red cells should be washed at least twice in normal saline before they
Solution IVD For in-vitro diagnostic use Use by/Expiration Date are finally washed and resuspended to 1.5-2% in LISS. This avoids
the non-specific uptake of autologous complement by the red cells
which can lead to unwanted positive reactions in anti-human globulin
REF: 284 001 4 x 10 ml tests.
REF: 284 002 4 x 100 ml
REF: 284 003 2 x 500 ml Direct agglutination tests at or below room temperature detect cold
antibodies, which are nearly always of no clinical significance, and
Intended Use consequently such techniques are not recommended for routine
LISS is used as potentiator for red blood cells, reducing the ionic antibody screening or compatibility testing. Unwanted positive
strength of the antibody: antigen reaction mixture by suspending monolayer of cells is indicative of a positive reaction. A compact reactions are less likely to be encountered if the temperature of the
red cells in LISS permits a substantial reduction in incubation time button of cells in the middle of the well is indicative of a negative red cell suspension, LISS or serum is in the range +19 ºC to +25ºC
and an increase in test sensitivity. These advantages of LISS are reaction. immediately before use.
entirely dependent on correct preparation and use. Therefore,
laboratories that use LISS techniques must take particular care Precautions and Warnings Red cells suspended in LISS should be clearly distinguished from red
with staff training. 1. In vitro diagnostic reagent for laboratory and professional use cells at normal ionic strength and should be discarded within 24 hours
only. Not for medicinal use. of preparation
Background 2. The reagent contains 0.09% sodium azide as a preservative.
In 1964 Mollison and Polley discovered that by reducing ion strength Avoid contamination with skin and mucosa. On disposal flush References
with a Low Ionic Strength Solution (LISS) antigen-antibody-reaction with large quantity of water. 1. Issitt, Peter D., and Issitt, Charla H. Applied Blood Group Serology
is considerably accelerated on blood group serological tests. 3. Do not freeze or expose the reagent to elevated temperatures. Oxnard, Calif.: Spectra Biologicals, 1979
LISS solution used as suspension medium for red cells and After usage immediately replace the bottle back to 2-8°C. 2. Walsh, RJ. “The effect of electrolytes on the Rh agglutination
contributing to reaction time reduction while at the same time 4. Turbidity may indicate reagent deterioration or contamination, reaction.” Med J Aust 1948;i:793.
increasing reaction strength. In antibody determination with such reagent should be discarded. 3. KJ Reis et al. Journal of Immunology 1984
coombsreactive antisera, LISS can be used in addition to the 5. Samples that cannot be tested immediately should be stored 4. Mark E. Brecher, MD et al. Technical Manual 15th Edition,
incubation medium. between +20 OC and +8 OC and tested within 48 hours. Bethesda.
Low Ionic Strength Salt Solutions (LISS) increase the rate of 6. All materials should be at room temperature before testing.
antigen-antibody complex formation. Additionally, since antibody 7. Do not use hemolysed or contaminated blood samples.
uptake is increased, incubation times of antigen-antibody reactions ORDERING INFORMATION
can be shortened. Excessive amounts of ions can interfere with Preparation, Storage & Stability CATALOG NO. QUANTITY
binding of antibody to antigen. Enhancement is achieved by Spectrum LISS is supplied ready-to-use and stable up to the
284 001 4 x 10 ml
decreasing the amount of ions in solution. expiry date labeled on the bottles when properly stored specified
284 002 4 x 100 ml
temperature. 284 003 2 x 500 ml
Assay Principle
Red blood cells are suspended in LISS Solution for cross match, Specimen Collection & Preparation
DAT, weak D typing and in LISS for antibody screening, identification For antibody screening, identification (Indirect Anti-
to be tested in the solid phase antiglobulin test Solidscreen II (please globulin Test IAT)
also refer to instructions for use of Solidscreen II).Solidscreen II is a Fresh samples of clotted or EDTA anticoagulated whole blood
solid phase assay for can be used for antibody screening, identification with the indirect
a) the detection of red blood cell alloantibodies or autoantibodies antiglobulin test. Samples collected following standard blood
in human plasma or serum. sampling guidelines are acceptable. The specimen should be
b) the determination of weak D and partial D antigens (DVI and tested as soon as possible after collection. If testing is delayed,
DVII) of donor samples which have tested negative with IgM EDTA and clotted specimens should be stored at 2 to 8°C. Use
anti-D using Erytype S and the. of samples older seven days should be avoided unless there is
The Solidscreen II well is coated with Protein A. Protein A is a no other alternative since antibody reactivity has been shown to
component of the cell wall of Staphylococcus aureus and has a decrease in older samples. Stored samples should be allowed to
very high affinity for the Fc portion of most immunoglobulin classes. reach room temperature prior to testing. Blood specimens exhibiting
For a) The plasma or serum and red blood cells (RBC, donor or gross hemolysis or contamination should not be used.
patient red blood cells) are added to the Protein-A coated well. There must be a distinct separation between the cellular and the
Sensitization of the red cell occurs if the corresponding antibody is plasma layer in the sample tube. Samples can be centrifuged or
present for the antigen on the red cell. allowed to settle.
For b) Solidscreen II Anti-D Blend and donor red blood cells are
added to the Protein- A coated well. Sensitization of the red blood Procedure
cell occurs if D antigen is present on the red blood cell. LISS techniques offer increased test sensitivity with decreased
Following incubation, and two wash processes to remove unbound incubation time. However, the benefits of LISS are entirely
protein, Anti- Human Globulin Anti-IgG Solidscreen II is added to dependent on the correct performance of techniques. For optimum
the well. Following centrifugation, the well is evaluated. A smooth sensitivity the LISS indirect antiglobulin technique requires a
minimum incubation time of 15 minutes.
266 267
H-Dry chemistry
268 269
CMV IgG/IgM Rapid Test- SYMBOLS IN PRODUCT LABELLING Plasma
Cassette EC REP Authorised Representative

1. Collect blood specimen into a lavender, blue or green
top collection tube (containing EDTA, citrate or heparin,
(Serum / Plasma) respectively in Vacutainer® ) by veinpuncture.
2. Separate the plasma by centrifugation.
REF Catalogue Number for use 3. Carefully withdraw the plasma into new pre-labeled tube.
REF: 1188 001 30 test
Consult instructions for use Manufactured by c. In addition to the presence of C band, if T lines in the both
coated Serum panel are developed, the test indicates for the presence
INTENDED USE 1. Collect blood specimen into a red top collection tube (containing of IgG and IgM anti-CMV. The result is CMV positive,
The Spectrum CMV IgG/IgM Rapid Test is a lateral flow no anticoagulants in Vacutainer®) by veinpuncture. indicating for a current infection.
immunoassay for the simultaneous detection and differentiation of CMV antigen, forming a burgundy colored T band, indicating a CMV 2. Allow the blood to clot.
IgM and IgG Cytomegalovirus (CMV) in human serum or plasma. IgM positive test result. 3. Separate the serum by centrifugation.
It is intended to be used as a screening test and as an aid in the 4. Carefully withdraw the serum into a new pre-labeled tube.
diagnosis of infection with CMV. Any reactive specimen with IgG anti-CMV, if present in the specimen, will bind to the CMV
the Spectrum CMV IgG/IgM Rapid Test must be confirmed with conjugates. The immunocomplex is then captured by the anti- Test specimens as soon as possible after collecting. Store
alternative testing method(s). human IgG on the membrane on the right panel, forming a burgundy specimens at 2°C-8°C if not tested immediately.
colored T band, indicating a CMV IgG positive test result. Samples with positive results should be confirmed with
SUMMARY AND EXPLANATION OF THE TEST Store specimens at 2°C-8°C up to 5 days. The specimens should be alternative testing method(s) and clinical findings before a
Cytomegalovirus (CMV) infections are widespread and usually Absence of any T bands suggests a negative result. The test frozen at -20°C for longer storage. positive determination is made.
asymptomatic; however, the virus may persist as a latent or chronic contains an internal control (C band) which should exhibit a
infection. The relatively frequent incidence and severe disease in burgundy colored band of the immunocomplex of goat anti rabbit Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen 3. INVALID: If no C band is developed, the assay is invalid
newborns and immunosuppressed individuals clearly establishes IgG/rabbit IgG-gold conjugate regardless of the color development specimens to room temperature slowly and mix gently. Specimens regardless of any burgundy color in the T bands as indicated
this agent as an important human pathogen. CMV infection can be on any of the T bands. Otherwise, the test result is invalid and the containing visible particulate matter should be clarified by below. Repeat the assay with a new device.
classified as Congenital (Acquired before birth), Perinatal (Acquired specimen must be retested with another device. centrifugation before testing. Note: Invalid test result in one device does not disqualify the
at birth) and Postnatal (Acquired after birth). The prognosis for test result on the other device as long as the result on the
congenitally infected infants who are asymptomatic at birth must REAGENTS AND MATERIALS PROVIDED ASSAY PROCEDURE other device is valid.
be guarded. Ten to 25% may subsequently develop hearing loss. 1. Each kit contains 25 or 30 test devices, each sealed in a foil Step 1: Bring the specimen and test components to room
Five to 10% may exhibit various degrees of mental retardation pouch with 3 items inside: temperature if refrigerated or frozen. Mix the specimen well prior to
and central nervous system motor disorders. Surveys show the a. One cassette device. assay once thawed.
incidence of congenital CMV infection to be from 0.5 to 2.5 %. b. One pipette dropper. Step 2: When ready to test, open the pouch at the notch and
Consequently, a careful documentation of the long-term effects of c. One desiccant. remove device. Place the test device on a clean, flat surface.
intrauterine infection is important. Although the age at which CMV 2. 2 bottles of the Sample Diluent (5 mL each bottle) Step 3: Be sure to label the device with specimen’s ID number.
infection is acquired varies with socioeconomic condition, only 3. One package insert (instruction for use). Step 4: Fill the pipette dropper with the specimen.
about 10-15% children in the United States are seropositive. By the Holding the dropper vertically, dispense 1 drop (about 30-45 µL) of PERFORMANCE CHARACTERISTICS
age 35 however, about 50% of the population is seropositive. MATERIALS REQUIRED BUT NOT PROVIDED specimen into To be established
1. Clock or Timer the sample well on each panel making sure that there are no air
The majority of individuals contracting postnatal CMV infections bubbles. LIMITATIONS OF TEST
remain asymptomatic. A small percentage of individuals will WARNINGS AND PRECAUTIONS Then add 1 drop (about 35-50 µL) of Sample Diluent immediately to 1. The Assay Procedure and the Test Result Interpretation must
develop a negative heterophile-antibody infectious mononucleosis For in Vitro Diagnostic Use the sample well on each panel. be followed closely when testing the presence of antibodies to
syndrome. In immunocompromised patients CMV infections happen 1. This package insert must be read completely before performing CMV in serum or plasma from individual subjects. Failure to
frequently, often from reactivation of latent infection, and may life- the test. Failure to follow the insert gives inaccurate test results. follow the procedure may give inaccurate results.
threatening (2-4). 2. Do not open the sealed pouch, unless ready to conduct the 2. The Spectrum CMV IgG/IgM Rapid Test is limited to the
assay. qualitative detection of antibodies to CMV in human serum or
Antibody of the IgM class is produced during the first 2-3 weeks of 3. Do not use expired devices. plasma. The intensity of the test band does not correlate with
infection with CMV and exists only transiently in most patients (8,9). 4. Bring all reagents to room temperature (15°C-30°C) before antibody titer of the specimen.
Serologic procedures which measure the presence of IgM and use. 3. A negative result for an individual subject indicates absence
IgG antibodies help discriminate between primary and recurrent 5. Do not use the components in any other type of test kit as a of detectable CMV antibodies. However, a negative test result
infections since IgM antibodies are rarely found in recurrent substitute for the components in this kit. Step 5: Set up timer. does not preclude the possibility of exposure to or infection
infections. 6. Do not use hemolized blood specimen for testing. Step 6: Results can be read in 15 minutes. with CMV.
7. Wear protective clothing and disposable gloves while 4. A negative result can occur if the quantity of the anti-virus
The Spectrum CMV IgG/IgM Rapid allows the discrimination of IgG handling the kit reagents and clinical specimens. Wash hands Don’t read result after 15 minutes. To avoid confusion, discard antibodies present in the specimen is below the detection
and IgM antibody in one test within 15 minutes. The test is user thoroughly after performing the test. the test device after interpreting the result limits of the assay, or the antibodies that are detected are
friendly, without cumbersome laboratory equipment. 8. Users of this test should follow the US CDC Universal not present during the stage of disease in which a sample is
Precautions for prevention of transmission of HIV, HBV and INTERPRETATION OF ASSAY RESULT collected.
TEST PRINCIPLE other blood-borne pathogens. 1. NEGATIVE RESULT: If only the C band is present, the 5. The results obtained with this test should only be interpreted
The Spectrum CMV IgG/IgM Rapid Test is a lateral flow 9. Do not smoke, drink, or eat in areas where specimens or kit absence of any burgundy color in the both T bands indicates in conjunction with other diagnostic procedures and clinical
chromatographic immunoassay panel device. The test cassette reagents are being handled. that no anti-CMV IgG neither anti-CMV IgM are detected. The findings.
consists of a CMV IgM detection panel (The left panel) and a 10. Dispose of all specimens and materials used to perform the result is negative.
CMV IgG detection panel (the right panel). The IgM panel has 1) a test as biohazardous waste. REFERENCES
burgundy colored conjugate pad containing mouse anti-human IgM 11. Handle the Negative and Positive Control in the same manner 1. Chamesky MA, Ray CG, and Smith TF:Laboratory diagnosis
conjugated with colloid gold (IgM conjugates), 2) a nitrocellulose as patient specimens. of viral infections. Cumitech 15, American Society for
membrane strip containing a test band (T band) and a control band 12. The testing results should be read within 10 minutes after Microbiology, Washington, DC,1982.
(C band). The T band is pre-coated with recombinant CMV antigen, a specimen is applied to the sample well or sample pad of 2. Jordon MC: Latent infection and the elusive cytomegalovirus.
and the C band is pre-coated with goat anti-mouse IgG.The IgG the device. Read result after 10 minutes may give erroneous 2. POSITIVE RESULT: Rev. Infect. Dis. 5:205-215, 1983.
panel has 1) a burgundy colored conjugate pad containing CMV results. a. In addition to the presence of C band, if only T band is 3. Weller TH:Clinical spectrum of cytomegalovirus infection. In:
antigens conjugated with colloid gold (CMV conjugates) and rabbit 13. Do not perform the test in a room with strong air flow, ie. an developed on the left panel, indicates for the presence Nahmias AJ, Dowdle WR, and Schinazi RF, eds. The Human
IgG-gold conjugates, 2) a nitrocellulose membrane strip containing electric fan or strong air-conditioning. of IgM anti- CMV; the result is positive for CMV test. It Herpes Vireuses, an interdisciplinary perspective. Elsevier/
a test band (T band) and a control band (C band). The T band is pre- indicates for a fresh infection. North Holland Publishing Co., New York, pp. 20-30, 1980.
coated with anti-human G antibody, and the C band is pre-coated 4. Adler SP:Transfusion-associated cytomegalovirus infections.
REAGENT PREPARATION AND STORAGE INSTRUC- Rev. Infect. Dis. 5:977-993, 1983
with goat anti-rabbit IgG. TIONS 5. Melish ME and Hanshaw JB:Congenital cytomegalovirus
All reagents are ready to use as supplied. Store unused test devices infection: Development progress of infants defected by routine
unopened at 2°C-30°C. The positive and negative controls should screening. Am. J. Dis. Child. 126:190-194, 1973.
be kept at 2°C-8°C. If stored at 2°C-8°C, ensure that the test device 6. Stagno S. Tinker Mk, Elrod C, Fuccillo DA, Cloud G, and
is brought to room temperature before opening. The test device is b. In addition to the presence of C band, if only T band is O’Beime AJ:Immuno-globulin M antibodies detected by
stable through the expiration date printed on the sealed pouch. Do developed on the right panel, the test indicates for the enzyme-linked immunosorbent assay and radioimmunoassay
not freeze the kit or expose the kit over 30°C presence of IgG anti-CMV. The result is CMV positive, in the diagnosis of cytomegalovirus infections in pregnant
When an adequate volume of test specimen is dispensed into the
sample well (S) of the cassette, the specimen migrates by capillary indicating for a past infection women and newborn infants. J. Clin. Micro 21:930-935, 1985.
SPECIMEN COLLECTION AND HANDLING
action across the cassette. The IgM antibody to CMV, if present Consider any materials of human origin as infectious and handle
in the specimen will bind to the IgM conjugates on the left panel. them using standard biosafety procedures.
The immunocomplex is then captured on the membrane by the pre-
270 271
H. pylori Ab Test Device SYMBOLS IN PRODUCT LABELLING
Test specimens as soon as possible after collecting. Store
specimens at 2°C-8°C if not tested immediately. Store specimens
test indicates for the presence of antibodies to H. pylori in the
specimen. The result is positive.
(Serum / Plasma / Whole Blood) at 2°C- 8°C up to 5 days. The specimens should be frozen at -20°C
for longer storage.
REF: 1180 001 30 test
Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen
REF Catalogue Number for use specimens to room temperature slowly and mix gently. Specimens Samples with positive results should be confirmed with alternative
Consult instructions for use Manufactured by containing visible particulate matter should be clarified by testing method(s) and clinical findings before a positive
centrifugation before testing. Do not use samples demonstrating determination is made.
INTENDED USE gross lipemia, gross hemolysis or turbidity in order to avoid
The Spectrum H. pylori Ab Test Device is a sandwich lateral flow 2. Sample Diluent (1 bottle, 5 mL) interference on result interpretation.
chromatographic immunoassay for the qualitative detection of 3. One package insert (instruction for use).
antibodies (IgG, IgM, and IgA) anti- Helicobacter pylori (H. pylori) Blood
in human serum, plasma, whole blood. It is intended to be used as MATERIALS MAY BE REQUIRED AND NOT PROVIDED Drops of whole blood can be obtained by either finger tip puncture 3. INVALID: If no C band is developed, the assay is invalid
a screening test and as an aid in the diagnosis of infection with H. 1. Positive Control or veinpuncture. Do not use any hemolized blood for testing. regardless of color development on the T band as indicated
pylori. Any reactive specimen with the Spectrum H. pylori Ab Test 2. Negative Control below. Repeat the assay with a new device.
Device must be confirmed with alternative testing method(s) and Whole blood specimens should be stored in refrigeration (2°C -8°C)
clinical findings. MATERIALS REQUIRED BUT NOT PROVIDED if not tested immediately. The specimens must be tested within 24
1. Clock or Timer hours of collection.
SUMMARY AND EXPLANATION OF THE TEST 2. Lancing device for whole blood test
Helicobacter pylori is associated with a variety of gastrointestinal ASSAY PROCEDURE
diseases included non-ulcer dyspepsia, duodenal and gastric WARNINGS AND PRECAUTIONS Step 1: Bring the specimen and test components to room PERFORMANCE CHARACTERISTICS
ulcer and active, chronic gastritis1,2. The prevalence of H. pylori For in Vitro Diagnostic Use temperature if refrigerated or frozen. Mix the specimen well prior to Clinical Performance
infection could exceed 90% in patients with signs and symptoms of 1. This package insert must be read completely before performing assay once thawed. A total of 200 specimens from the non- H. pylori patients and 75
gastrointestinal diseases. Recent studies indicate an association of the test. Failure to follow the insert gives inaccurate test results. Step 2: When ready to test, open the pouch at the notch and specimens from the patients under anti-H. pylori treatment were
H. pylori infection with stomach cancer3. 2. Do not open the sealed pouch, unless ready to conduct the remove device. Place the test device on a clean, flat surface. tested by the Spectrum H. pylori Ab Test Device. Comparison for all
assay. Step 3: Be sure to label the device with specimen’s ID number. subjects is shown in the following table.
H. pylori colonizing in the gastrointestinal system elicits specific 3. Do not use expired devices. Step 4: Fill the plastic dropper with the specimen.
Spectrum H. pylori Ab Test Device
antibody responses4,5,6 which aids in the diagnosis of H. pylori 4. Bring all reagents to room temperature (15°C-30°C) before Holding the dropper vertically, dispense 1 drop (about 30-45 µL) of
infection and in monitoring the prognosis of the treatment of H. use. serum/plasma or 1 drop of whole blood (about 40-50 µL) into the H. pylori Patients Positive Negative Total
pylori related diseases. Antibiotics in combination with bismuth 5. Do not use the components in any other type of test kit as a sample well making sure that there are no air bubbles.
Positive 65 10 75
compounds have been shown to be effective in treating active H. substitute for the components in this kit. Immediately add 1 drop (about 35-50 µL) of Sample Diluent to the
pylori infection. Successful eradication of H. pylori is associated sample well. Negative 18 182 200
6. Do not use hemolized blood specimen for testing.
with clinical improvement in patients with gastrointestinal diseases 7. Wear protective clothing and disposable gloves while Total 83 180 275
providing a further evidence7. handling the kit reagents and clinical specimens. Wash hands
thoroughly after performing the test. Relative Sensitivity: 86.7% , Relative Specificity: 91%, Overall
The Spectrum H. pylori Ab Test Device is a latest generation of 8. Users of this test should follow the US CDC Universal Agreement: 89.8%
chromatographic immunoassay which utilizes recombinant antigens Precautions for prevention of transmission of HIV, HBV and
to detect the antibodies to H. pylori in human serum or plasma. The other blood-borne pathogens. LIMITATIONS OF TEST
test is user friendly, highly sensitive and specific. 9. Do not smoke, drink, or eat in areas where specimens or kit 1. The Assay Procedure and the Assay Result Interpretation must
reagents are being handled. be followed closely when testing the presence of antibodies
TEST PRINCIPLE 10. Dispose of all specimens and materials used to perform the to H. pylori in serum, plasma or whole blood from individual
The Spectrum H. pylori Ab Test Device is a lateral flow test as biohazardous waste. subjects. Failure to follow the procedure may give inaccurate
11. Handle the Negative and Positive Control in the same manner Step 5: Set up timer.
chromatographic immunoassay based on the principle of the results.
as patient specimens. Step 6: Results can be read in 15 minutes. Positive results can
double antigen–sandwich technique. The test cassette consists of: 2. The Spectrum H. pylori Ab Test Device is limited to the
12. The testing results should be read within 15 minutes after be visible in as short as 1 minute.
1) a burgundy colored conjugate pad containing H. pylori antigens qualitative detection of IgG, IgM, and IgA anti- H. pylori in
including Cag-A conjugated with colloid gold (H. pylori conjugates) a specimen is applied to the sample well or sample pad of human serum, plasma or whole blood. The intensity of the test
the device. Read result after 15 minutes may give erroneous Don’t read result after 15 minutes. To avoid confusion, discard
and rabbit IgG-gold conjugates, 2) a nitrocellulose membrane strip band does not have linear correlation with the antibody titer in
results. the test device after interpreting the result.
containing a test band (T band) and a control band (C band). The T the specimen.
band is pre-coated with non-conjugated H. pylori antigens, and the 13. Do not perform the test in a room with strong air flow, ie. 3. A negative result for an individual subject indicates absence
C band is pre-coated with goat anti-rabbit IgG. electric fan or strong air-conditioning. QUALITY CONTROL
of detectable antibodies to H. pylori. However, a negative
1. Internal Control: This test contains a built-in control feature,
test result does not preclude the possibility of exposure to or
REAGENT PREPARATION AND STORAGE the C band. The C line develops after adding specimen and
infection with H. pylori.
sample diluent. Otherwise, review the whole procedure and
INSTRUCTIONS 4. A negative result can occur if the quantity of the antibodies to
repeat test with a new device.
All reagents are ready to use as supplied. Store unused test device H. pylori present in the specimen is below the detection limits
2. External Control: Good Laboratory Practice recommends
unopened at 2°C-30°C. If stored at 2°C-8°C, ensure that the test of the assay, or the antibodies that are detected are not present
using the external controls, positive and negative, to assure
device is brought to room temperature before opening. The test during the stage of disease in which a sample is collected.
the proper performance of the assay, particularly under the
device is stable through the expiration date printed on the sealed 5. Some specimens containing unusually high titer of heterophile
following circumstances:
When an adequate volume of test specimen is dispensed into the pouch. Do not freeze the kit or expose the kit over 30°C. antibodies or rheumatoid factor may affect expected results.
a. New operator uses the kit, prior to performing testing of
sample well of the cassette, the specimen migrates by capillary 6. The results obtained with this test should only be interpreted
specimens.
action across the cassette. The antibodies: either the IgG, the IgM, SPECIMEN COLLECTION AND HANDLING in conjunction with other diagnostic procedures and clinical
b. A new lot of test kit is used.
or the IgA, to H. pylori if present in the specimen will bind to the Consider any materials of human origin as infectious and handle findings.
c. A new shipment of kits is used.
H. pylori conjugates. The immunocomplex is then captured on the them using standard biosafety procedures. d. d. The temperature used during storage of the kit falls
membrane by the pre-coated H. pylori antigens, forming a burgundy Plasma REFERENCES
outside of 2-30OC.
colored T band, indicating a H. pylori Ab positive test result. 1. Collect blood specimen into a lavender, blue or green 1. Megraud,F.et.al.1989. 27:1870-3,1989
e. The temperature of the test area falls outside of 15
top collection tube (containing EDTA, citrate or heparin, - 30OC. 2. Parsonnet,J.et.al.1991. New England J. Med. 325:1127-31.
Absence of the T band suggests a negative result. The test contains respectively in Vacutainer® ) by veinpuncture. 3. Marshall,B.J.et.al.1985. Med. J. Australia. 149:439-44,
f. To verify a higher than expected frequency of positive or
an internal control (C band) which should exhibit a burgundy 2. Separate the plasma by centrifugation. 4. Ansong,R. et.al.1991. J.Clin.Micro. 29:51-53,
negative results.
colored band of the immunocomplex of goat anti-rabbit IgG/rabbit 3. Carefully withdraw the plasma into new pre-labeled tube. 5. Soll,A.H. 1990. New England J. Med.322:909-916.
g. To investigate the cause of repeated invalid results.
IgG-gold conjugate regardless the presence of any antibodies to H. 6. Pronovost,A.P.et.al. 1994. J.Clin.Microbiol.32:46-50.
pylori. Otherwise, the test result is invalid and the specimen must be Serum
retested with another device. 1. Collect blood specimen into a red top collection tube (containing INTERPRETATION OF ASSAY RESULT
no anticoagulants in Vacutainer®) by veinpuncture. 1. NEGATIVE RESULT: If only the C band is developed, the test
REAGENTS AND MATERIALS PROVIDED 2. Allow the blood to clot. indicates that no detectable antibodies to H. pylori are present
1. Individually sealed foil pouches containing: 3. Separate the serum by centrifugation. in the specimen. The result is negative.
a. One cassette device. 4. Carefully withdraw the serum into a new pre-labeled tube. 2. POSITIVE RESULT: If both C and T bands are developed, the
b. One plastic dropper.
c. One desiccant.
272 273
H. pylori Ag Test Device SYMBOLS IN PRODUCT LABELLING
regardless of color development on the T band as indicated
below. Repeat the assay with a new device. If it is caused
(Fecal Specimen) EC REP Authorised Representative Temperature Limitation by excess amount of fecal specimen collected, re-sample
and re-test.
REF: 1184 001 30 test
The specimen is now ready for testing, transportation or storage.
REAGENTS AND MATERIALS PROVIDED
1. Individually sealed foil pouches containing:
Note: Specimens collected may be stored 3 days at 2°C -8°C, PERFORMANCE CHARACTERISTICS
or 1 year at <-20°C Clinical Performance
INTENDED USE a. One cassette test device.
328 fecal samples collected from subjects with symptomatic
The Spectrum H. pylori Ag Test Device is a lateral flow b. One desiccant.
2. Sample extraction tubes, each containing 2 mL of extraction
TEST PROCEDURE gastrointestinal disorders and non-gastrointestinal symptoms were
chromatographic immunoassay for the qualitative detection of H.
Step 1: Bring the specimen and test components to room tested with the Spectrum H. pylori Ag Test Device with the UBT
pylori antigen in human fecal specimen. It is intended to be used by buffer.
temperature if refrigerated or frozen. as reference test. Comparison for all subjects is shown in the
professionals as a screening test and as an aid in the diagnosis of 3. One package insert (instruction for use).
Step 2: When ready to test, open the pouch at the notch and following table:
infection with H. pylori. Any reactive specimen with the Spectrum
remove the test strip. Place the test strip on a clean, flat surface.
H. pylori Ag Test Device must be confirmed with alternative testing MATERIALS MAY BE REQUIRED AND AVAILABLE FOR Spectrum H. pylori Ag Test Device
Step 3: Shake the sample collection tube vigorously to ensure an
method(s) and clinical findings. PURCHASE
effective liquid suspension. UBT Positive Negative Total
1. Positivia H. pylori Ag Test Device Assay Control Kit (Cat #
Step 4: Hold the tube upright, twist off the tip. Dispense 2 drops
SUMMARY AND EXPLANATION OF THE TEST C0192) contains one vial of positive control and one vial of
of the solution into the sample pad (s) of the strip. Do not over load Positive 118 7 125
Helicobacter pylori is associated with a variety of gastrointestinal negative control
samples. Negative 0 199 199
diseases included non-ulcer dyspepsia, duodenal and gastric
ulcer and active, chronic gastritis1,2. The prevalence of H. pylori MATERIALS REQUIRED BUT NOT PROVIDED Total 118 206 324
infection could exceed 90% in patients with signs and symptoms of 1. Clock or Timer
gastrointestinal diseases. Recent studies indicate an association of 2. A container to hold fecal specimen Relative Sensitivity: 94.4% , Relative Specificity: 100.0%, Overall
H. pylori infection with stomach cancer3. Agreement: 97.8%
WARNINGS AND PRECAUTIONS Analytic Sensitivity:
H. pylori can be transmitted by means of oral–fecal matter For in Vitro Diagnostic Use The detection limit for the Spectrum H. pylori Ag Test Device -is 5
through the ingestion of waste tainted food or water. Antibiotics in 1. This package insert must be read completely before performing ng/ml of H. pylori lysate. The fecal specimen extraction contains H.
combination with bismuth compounds showed to be effective in the test. Failure to follow the insert gives inaccurate test pylori lysate equal to or greater than 5 ng/ml routinely test positive.
treating active H. pylori infection. results. Specimens containing H. pylori lysate less than 5 ng/ml may also
2. Do not open the sealed pouch, unless ready to conduct the produce a very faint positive line, especially with extended assay
Step 5: Set up the timer.
H. pylori infection is currently detected by invasive testing methods assay. time beyond 15 minutes.
Step 6: Results can be read in 15 minutes after adding the
(ie histology, culture) based on endoscopy and biopsy, or non- 3. Do not use any kit components beyond their stated expiration The following experiments were done to validate the sensitivity of
specimen. Positive results can be visible in as short as 1 minute.
invasive testing methods, such as urea breath test (UBT), serologic date. the Spectrum H. pylori Ag Test Device -Card:
Don’t read results after 15 minutes. To avoid confusion, dis-
antibody test and stool antigen test. UBT requires one month long 4. Do not use the components in any other type of test kit as a Normal fecal extraction were spiked with H. pylori lysate to
card the test device after interpreting the result.
preparation and consume radioactive material. Serologic antibody substitute for the components in this kit. concentrations of 0, 1.25, 2,5, 5, 10, 20 ng/ml. The specimens
tests do not distinguish between currently active infection with a past 5. Bring all reagents to room temperature (15°C-30°C) before were run on the Spectrum H. pylori Ag Test Device -. Results are
use.
QUALITY CONTROL
exposure or an infection that has been cured. The stool antigen tabulated in table below.
1. Internal Control: This test contains a built-in control feature,
test detects antigen presence in the feces that indicates active H. 6. Wear protective clothing and disposable gloves while
the C band. The C line develops after adding specimen H. pylori lysate ng/ml 0 1.25 2.5 5 10 20
pylori infection. It can be also used to monitor the effectiveness of handling the kit reagents and clinical specimens. Wash hands
extract. Otherwise, review the whole procedure and repeat
treatment and the recurrence of the infection. thoroughly after performing the test. Number of positive 0 0 12 20 20 20
test with a new device.
7. Users of this test should follow the US CDC Universal
2. External Control: Good Laboratory Practice recommends Number of negative 20 20 8 0 0 0
The Spectrum H. pylori Ag Test Device uses a colloid gold conjugated Precautions for prevention of transmission of HIV, HBV and
using the external controls, positive and negative, to assure
monoclonal anti- H. pylori antibody and another monoclonal anti-H. other blood-borne pathogens.
the proper performance of the assay, particularly under the n=20 relative sensitivity at 5 ng/ml = 20/20 x 100% = 100%
pylori antibody to specifically detect H. pylori antigen present in the 8. Do not smoke, drink, or eat in areas where specimens or kit
following circumstances:
infected patient fecal specimen. The test is user friendly, accurate, reagents are being handled.
a. New operator uses the kit, prior to performing testing of LIMITATIONS OF TEST
and the result is available instantly. 9. Extraction buffer contains 0.1% NaN3. Avoid contact with skin
specimens. 1. The Assay Procedure and the Test Result Interpretation must
or eyes. Do not ingest.
b. A new lot of test kit is used. be followed closely when testing the presence of H. pylori
TEST PRINCIPLE 10. Dispose of all specimens and materials used to perform the
c. A new shipment of kits is used.
The Spectrum H. pylori Ag Test Device is a sandwich lateral flow test as biohazardous waste. antigen in feces. Failure to follow the procedure, in particularly
d. The temperature used during storage of the kit falls sampling procedure, may give inaccurate results.
chromatographic immunoassay. The test strip consists of: 1) a 11. The testing results should be read within 15 minutes after a
outside of 2-30OC. 2. The Spectrum H. pylori Ag Test Device is limited to the
burgundy colored conjugate pad containing monoclonal anti-H. specimen is applied to the sample well of the device. Read
e. The temperature of the test area falls outside of 15 -30OC. qualitative detection of H. pylori antigen in human fecal
pylori antibody conjugated with colloid gold (anti-H.P conjugates) result after 15 minutes may give erroneous results.
f. To verify a higher than expected frequency of positive or specimen. The intensity of the test band does not have linear
and 2) a nitrocellulose membrane strip containing a test band (T 12. Do not perform the test in a room with strong air flow, ie.
negative results. correlation with antigen title in the specimen.
band) and a control band (C band). The T band is pre-coated with electric fan or strong air-conditioning.
g. To investigate the cause of repeated invalid results. 3. A negative result for an individual subject indicates absence of
another monoclonal anti-H.P antibody, and the C band is pre-coated
with goat anti-mouse IgG antibody. REAGENT PREPARATION AND STORAGE detectable H. pylori. Antigen. However, a negative test result
INSTRUCTIONS INTERPRETATION OF ASSAY RESULT does not preclude the possibility of infection with H. pylori.
1. NEGATIVE RESULT: If only the C band is developed, the test 4. A negative result can occur if the quantity of the H. pylori
All reagents are ready to use as supplied. Store unused test devices
indicates that no detectable H. pylori antigen is present in the antigen present in the specimen is below the detection limits of
unopened at 2°C-30°C. If stored at 2°C-8°C, ensure that the test
specimen. The result is negative. the assay, or the antigen that are detected are not present in
device is brought to room temperature before opening. The test
device is stable through the expiration date printed on the sealed fecal sample is collected.
pouch. Do not freeze the kit or expose the kit over 30°C. 5. If the symptom persists, while the result from Spectrum H.
pylori Ag Test Device is negative or non-reactive result, it is
When an adequate volume of extracted fecal specimen is dispensed SPECIMEN COLLECTION AND HANDLING recommended to re-sample the patient few days late or test
into the sample well of the test cassette, the specimen migrates by Consider any materials of human origin as infectious and handle with an alternative test device.
2. POSITIVE RESULT: If both C and T bands are developed, 6. The results obtained with this test should only be interpreted
capillary action across the cassette. H.P antigen if present in the them using standard biosafety procedures.
the test indicates for the presence of H. pylori antigen in the in conjunction with other diagnostic procedures and clinical
specimen will bind to the anti-H.P conjugates. The immunocomplex 1. Collect a random sample of feces in a clean, dry receptacle.
specimen. The result is positive. findings.
is then captured on the membrane by the pre-coated antibody, 2. Unscrew the top of the collection tube and remove the
forming a burgundy colored T band, indicating a H.P positive test applicator stick.
result. Absence of this band suggests that the concentration of 3. Randomly pierce the fecal specimen in at least five (5) different REFERENCES
H.P in the specimen is below the detectable level, indicating a H.P sites. Do not scoop fecal specimen as this will lead to 1. Vans DJ, Evans DG, et at A sensitive and specific seriologic
negative result. invalid test result. test for detection of campylobacter pylori infection.
4. Remove excess sample off the shaft and outer grooves. Be Gastroenerology. 1989, 96:1004
Samples with positive results should be interpreted in conjunction 2. Lambert IR, Lin SK, and Aranda-Michel. J, helocobacter pylori
Absence of the T band suggests a negative result. The test contains sure specimen remains on inside grooves. Specimen on the
with other testing procedure and clinical findings before a Scan. J. Gasteroenterol. 1995, 30 suppl 208: 33-46
an internal control (C band) which should exhibit a burgundy grooves is sufficient for testing. Excess amount of fecal can
diagnostic decision is made. 3. Marshall BJ, et al, pyloric camplylobacter infection and
colored band of the immunocomplex of goat anti-mouse IgG/mouse lead to invalid test result.
IgG-gold conjugate regardless of the color development on the T 5. Replace the stick in the tube and tighten securely. gastroduodenal disease. Med.J. Aust. 1985, 142: 439-444.
3. INVALID: If no C band is developed, the assay is invalid
band. Otherwise, the test result is invalid and the specimen must be
retested with another device.
274 275
HAV IgM Rapid Test-Cassette SYMBOLS IN PRODUCT LABELLING
be frozen at -20°C for longer storage.
Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen
Samples with positive results should be confirmed with alternative
testing method(s) and clinical findings before a positive
(Serum / Plasma) EC REP Authorised Representative Temperature Limitation specimens to room temperature slowly and mix gently. Specimens determination is made.
IVD For in-vitro diagnostic use Use by/Expiration Date containing visible particulate matter should be clarified by
REF: 1192 001 30 test centrifugation before testing. 3. INVALID: If no C line is developed, the assay is invalid
regardless of color development on the T line as indicated
ASSAY PROCEDURE below. Repeat the assay with a new device.
Step 1: Bring the specimen and test components to room
temperature if refrigerated or frozen. Mix the specimen well, prior to
INTENDED USE assay, once thawed.
The Spectrum HAV IgM Rapid Test is a lateral flow chromatographic
immunoassay for the qualitative detection of IgM antibodies to MATERIALS REQUIRED BUT NOT PROVIDED Step 2: When ready to test, open the pouch at the notch and
Hepatitis A virus (HAV) in human serum or plasma. It is intended 1. Clock or Timer remove device. Place the test device on a clean, flat surface.
to be used as a screening test and as an aid in the diagnosis of PERFORMANCE CHARACTERISTICS
infection with HAV. Any reactive specimen with the Spectrum HAV WARNINGS AND PRECAUTIONS Step 3: Be sure to label the device with the specimen’s ID Clinical Performance
IgM Rapid Test must be confirmed with alternative testing method(s) For in Vitro Diagnostic Use number. A total of 200 samples from susceptible subjects were tested by
and clinical findings. 1. This package insert must be read completely before performing the Spectrum HAV IgM Rapid Test and by a commercial EIA test.
the test. Failure to follow the insert gives inaccurate test results. Step 4: Fill the plastic dropper with the specimen. Comparison of the results for all subjects is shown in the following
SUMMARY AND EXPLANATION OF THE TEST 2. Do not open the sealed pouch unless ready to conduct the Holding the dropper vertically, dispense 1 drop (about 30-45 µL) t a b l e :
HAV is a positive-sense RNA virus, a unique member of assay. of specimen into the sample well making sure that there are no air
Spectrum HAV IgM Rapid Test
picornaviridae1. Its transmission depends primarily on serial 3. Do not use expired devices. bubbles.
transmission from person to person by the fecal-oral route. Although 4. Bring all reagents to room temperature (15°C-30°C) before Then add 1 drop (about 35-50 µL) of Sample Diluent immediately. EIA Positive Negative Total
hepatitis A is not ordinarily a sexually transmitted disease, the use. Positive 21 1 22
infection rate is high among male homosexuals as result of oral- 5. Do not use the components in any other type of test kit as a
anal contact. substitute for the components in this kit. Negative 0 178 178
6. Do not use hemolized blood specimens for testing. Total 21 179 200
The presence of specific anti-HAV IgM in blood samples suggests 7. Wear protective clothing and disposable gloves while
acute or recent HAV infection4-6. The IgM antibody rapidly increases handling the kit reagents and clinical specimens. Wash hands Relative Sensitivity: 95.5% , Relative Specificity: 100%, Overall
in titer over a period of 4-6 weeks post infection and then declines to thoroughly after performing the test. Agreement: 99.5%
non-detectable levels within 3 to 6 months in most patients7. 8. Users of this test should follow the US CDC Universal
Precautions for prevention of transmission of HIV, HBV and
other blood-borne pathogens.
LIMITATIONS OF TEST
The Spectrum HAV IgM Rapid Test is to be used to detect IgM
Step 5: Set up timer. 1. The Assay Procedure and the Interpretation of Assay Result
anti-HAV in less than 15 minutes by untrained or minimally skilled 9. Do not smoke, drink or eat in areas where specimens or kit
sections must be followed closely when testing for the
personnel, without cumbersome laboratory equipment. reagents are being handled.
Step 6: Results can be read in 15 minutes. Positive results can presence of anti-HAV IgM in serum or plasma from individual
10. Dispose of all specimens and materials used to perform the
be visible in as short as 1 minute. subjects. Failure to follow the procedure may give inaccurate
TEST PRINCIPLE test as bio-hazardous waste.
results.
The Spectrum HAV IgM Rapid Test is a lateral flow chromatographic 11. Handle the negative and positive controls in the same manner
Don’t read result after 15 minutes. To avoid confusion, discard 2. The Spectrum HAV IgM Rapid Test is limited to the qualitative
immunoassay. The test cassette consists of: 1) a burgundy as patient specimens.
the test device after interpreting the result. detection of anti-HAV IgM in human serum or plasma. The
colored conjugate pad containing mouse anti-human IgM 12. The testing results should be read within 15 minutes after
intensity of the test line does not have linear correlation with
antibody conjugated with colloidal gold (IgM conjugates) and 2) a a specimen is applied to the sample well or sample pad of
the antibody titer in the specimen.
nitrocellulose membrane strip containing a test band (T band) and a the device. Reading the results after 15 minutes may give QUALITY CONTROL 3. A negative result for an individual subject indicates absence
control band (C band). The T band is pre-coated with recombinant erroneous results. 1. Internal Control: This test contains a built-in control feature,
of detectable anti-HAV IgM. However, a negative test result
HAV antigen, and the C band is pre-coated with goat anti-mouse 13. Do not perform the test in a room with strong air flow, i.e. the C line. The C line develops after adding the specimen and
does not preclude the possibility of exposure to or infection
IgM antibodies. electric fan or strong air-conditioning. the sample diluent. If the C line does not develop, review the
with HAV.
whole procedure and repeat the test with a new device.
4. A negative result can occur if the quantity of the anti-HAV
REAGENT PREPARATION AND STORAGE 2. External Control: Good Laboratory Practice recommends
IgM present in the specimen is below the detection limits of
INSTRUCTIONS using external controls, positive and negative, to assure
the assay or the antibodies that are detected are not present
All reagents are ready to use as supplied. Store unused test device the proper performance of the assay, particularly under the
during the stage of disease in which a sample is collected.
unopened at 2°C-30°C. Do not expose the kit over 30°C. Do not following circumstances:
5. Some specimens containing unusually high titers of heterophile
freeze the kit. The negative and positive controls should be store a. New operator uses the kit, prior to performing the testing
antibodies or rheumatoid factor may affect expected results.
at 2-8°C or the temperature indicated. If stored at 2°C-8°C, ensure of specimens.
6. The results obtained with this test should only be interpreted
When an adequate volume of test specimen is dispensed into the that the test device is brought to room temperature before opening. b. A new lot of test kits is used.
in conjunction with other diagnostic procedures and clinical
sample well of the cassette, the specimen migrates by capillary The test device is stable through the expiration date printed on the c. A new shipment of kits is used.
findings.
action across the cassette. Anti-HAV IgM if present in the specimen sealed pouch. d. The temperature used during storage of the kits fall
will bind to the IgM conjugates. The immunocomplex is then outside of 2-30°C.
e. The temperature of the test area falls outside of 15-30°C. REFERENCES
captured on the membrane by the pre-coated HAV antigen forming SPECIMEN COLLECTION AND HANDLING
f. To verify a higher than expected frequency of positive or 1. Minor P. Picornaviridae. In: Francki RIB, Fauquet CM, Knudson
a burgundy colored T line, indicating a HAV IgM positive test result. Consider any materials of human origin as infectious and handle
negative results. DL, et al., eds. Classification and nomenclature of viruses
Absence of the T line suggests a negative result. them using standard bio-safety procedures.
g. To investigate the cause of repeated invalid results. (Arch Virol Supp 2). Wien: Springer-Verlag, 1991: 320-326.
Plasma 2. Keeffe EB. Clinical approach to viral hepatitis in homosexual
The test contains an internal control (C line) which should exhibit 1. Collect blood specimen into a lavender, blue or green men. Med Clin North Am. 1986;70(3):567-86.
a burgundy colored line of the immunocomplex of goat anti-mouse top collection tube (containing EDTA, citrate or heparin, INTERPRETATION OF ASSAY RESULT 3. Ballesteros J, Dal-Re R, Gonzalez A, del Romero J. Are
IgG/IgM-gold conjugate regardless of the color development of the respectively in Vacutainer® ) by veinpuncture. 1. NEGATIVE RESULT: If only the C line is developed, the test
homosexual males a risk group for hepatitis A infection in
T line. Otherwise, the test result is invalid and the specimen must be 2. Separate the plasma by centrifugation. indicates that no detectable IgM anti-HAV is present in the
intermediate endemicity areas? Epidemiol Infect. 1996;
retested with another device. 3. Carefully withdraw the plasma into new pre-labeled tube. specimen. The result is negative.
117(1):145-8.
4. Decker RH, Kosakowski SM, Vanderbilt AS, et al: Diagnosis of
REAGENTS AND MATERIALS PROVIDED Serum acute hepatitis A by HAVAB-M : A direct radioimmunoassay for
1. Individually sealed foil pouches containing:: 1. Collect blood specimen into a red top collection tube IgM anti-HAV. Am J Clin Pathol 1981;76:140-147.
a. One cassette device (containing no anticoagulants in Vacutainer®) by veinpuncture. 5. Locarnini SA, Ferris AA, Lehman NI, et al: The antibody
b. One desiccant 2. Allow the blood to clot. response following hepatitis A infection. Intervirology 1974;
2. Plastic droppers 3. Separate the serum by centrifugation. 2. POSITIVE RESULT: If both the C and the T lines are
4:110-118.
3. Sample Diluent (1 vial, 5 mL) 4. Carefully withdraw the serum into a new pre-labeled tube. developed, the test indicates the presence of IgM anti-HAV in
6. Skinhoj P, Mikkelsen F, Hollinger FB. Hepatitis A in Greenland:
4. One package insert (instruction for use) the specimen. The result is positive.
Importance of specific antibody testing in epidemiologic
Test specimens as soon as possible after collecting. Store surveillance. Am J. Epidemiol 1977; 105: 104-147.
MATERIALS MAY BE REQUIRED AND NOT PROVIDED specimens at 2°C-8°C if not tested immediately.
1. Positive Control
2. Negative Control Store specimens at 2°C-8°C for up to 5 days. The specimens should
276 277
HBsAg SYMBOLS IN PRODUCT LABELLING
INTERPRETATION OF RESULTS
(Please refer to the illustration above)
Specificity
Antibodies used for the HBsAg One Step Hepatitis B Surface
Hepatitis B Surface Antigen Test Device EC REP Authorised Representative Temperature Limitation POSITIVE: *Two distinct red lines appear. One line should be in the Antigen Test Device (Serum/Plasma) were developed against
(Serum/Plasma) IVD For in-vitro diagnostic use Use by/Expiration Date control region (C) and another line should be in the test region (T). whole Hepatitis B antigen isolated from Hepatitis B virus. Specificity
LOT Batch Code/Lot number CAUTION. Consult *NOTE: The intensity of the red color in the test line region (T) of the HBsAg One Step Hepatitis B Surface Antigen Test Device
REF: 1160 001 25 test will vary depending on the concentration of HBsAg present in the (Serum/Plasma) was also tested with laboratory strains of Hepatitis
REF Catalogue Number instructions for use
REF: 1160 002 50 test specimen. Therefore, any shade of red in the test region (T) should A and Hepatitis C. They all yielded negative results.
be considered positive.
A rapid, one step test for the qualitative detection of Hepatitis B NEGATIVE: One red line appears in the control region (C). No HBsAg Reference Method
Surface Antigen (HBsAg) in serum or plasma. apparent red or pink line appears in the test region (T). Method EIA Total
For professional in vitro diagnostic use only. INVALID: Control line fails to appear. Insufficient specimen volume
Results Positive Negative Results
or incorrect procedural techniques are the most likely reasons for HBsAg
INTENDED USE SPECIMEN COLLECTION AND PREPARATION ontrol line failure. Review the procedure and repeat the test with a Test Positive 145 5 150
The HBsAg One Step Hepatitis B Surface Antigen Test Device • The HBsAg One Step Hepatitis B Surface Antigen Test Device new test device. If the problem persists, discontinue using the test Device
(Serum/Plasma) is a rapid chromatographic immunoassay for the kit immediately and contact your local distributor. Negative 0 150 150
(Serum/Plasma) can be performed using either serum or
qualitative detection of Hepatitis B Surface Antigen in serum or plasma. Total Results 145 155 300
plasma. • Separate the serum or plasma from blood as soon as possible QUALITY CONTROL
to avoid hemolysis. Only clear, non-hemolyzed specimens can Relative Sensitivity: > 99.0%
SUMMARY Internal Quality Control Relative Specificity: 96.7%
be used.
Viral hepatitis is a systemic disease primarily involving the liver. • Testing should be performed immediately after the specimens A procedural control is included in the test. A colored line appearing Accuracy: 98.3%
Most cases of acute viral hepatitis are caused by Hepatitis A virus, have been collected. Do not leave the specimens at room in the control line region (C) is considered an internal procedural
control. It confirms sufficient specimen volume, adequate membrane Precision
Hepatitis B virus (HBV) or Hepatitis C virus. The complex antigen temperature for prolonged periods. Specimens may be stored
wicking and correct procedural technique. Intra-Assay
found on the surface of HBV is called HBsAg. Previous designations at 2-8°C for up to 3 days. For long term storage, specimens
included the Australia or Au antigen .1 The presence of HBsAg in Within-run precision has been determined by using 15 replicates
should be kept below - 20°C. External Quality Control
serum or plasma is an indication of an active Hepatitis B infection, of three specimens containing 0 ng/mL, 1 ng/mL and 5 ng/mL of
• Bring specimens to room temperature prior to testing. Frozen It is recommended that a positive and negative external control be
either acute or chronic. In a typical Hepatitis B infection, HBsAg will HBsAg. The negative and positive values were correctly identified
specimens must be completely thawed and mixed well prior run every 20 tests, and as deemed necessary by internal laboratory
be detected 2 to 4 98% of the time.
to testing. Specimens should not be frozen and thawed procedures. Some commercial controls may contain interfering
weeks before the ALT level becomes abnormal and 3 to 5 weeks repeatedly. preservatives; therefore, interpretation of results with external Inter-Assay
before symptoms or jaundice develop. HBsAg has four principal • If specimens are to be shipped, they should be packed in quality controls should be done with caution. Between-run precision has been determined by using the same
subtypes: adw, ayw, adr and ayr. Because of antigenic heterogeneity compliance with federal, state or local regulations for the three specimens of 0 ng/mL, 1ng/mL and 5 ng/mL of HBsAg in 15
of the determinant, there are 10 major serotypes of Hepatitis B virus. transportation of etiologic agents. Procedure for External Quality Control Testing independent assays. Three different lots of the HBsAg One Step
The HBsAg One Step Hepatitis B Surface Antigen Test Device 1. Hold the Control vial vertically and add 2 full drops of Control Hepatitis B Surface Antigen Test Device (Serum/Plasma) has been
(Serum/Plasma) is a rapid test to qualitatively detect the presence to the Specimen well (S) as in procedure Step 2. tested over a 3-month period using negative, low positive and high
of HBsAg in serum or plasma specimen. The test utilizes a MATERIALS 2. Continue with Step 3 of Directions For Use. If the controls positive specimens. The specimens were correctly identified 98%
combination of monoclonal and polyclonal antibodies to selectively Materials Provided do not yield the expected results, do not use the test results. of the time.
detect elevated levels of HBsAg in serum or plasma. • Test devices Repeat the test or contact your distributor.
• Disposable specimen droppers
• Package insert BIBLIOGRAPHY
PRINCIPLE LIMITATION 1. Blumberg, B.S. The Discovery of Australian Antigen and its
The HBsAg One Step Hepatitis B Surface Antigen Test Device Materials Required But Not Provided 1. The HBsAg One Step Hepatitis B Surface Antigen Test Device relation to viral hepatitis. Vitro. 1971; 7: 223
(Serum/Plasma) is a qualitative,lateral flow immunoassay for the • Specimen collection containers (Serum/Plasma) is for in vitro diagnostic use only. This test
detection of HBsAg in serum or plasma. The membrane is pre- • Centrifuge should be used for the detection of HBsAg in serum or plasma
coated with anti-HBsAg antibodies on the test line region of the • Timer specimen.
test. During testing, the serum or plasma specimen reacts with the 2. The HBsAg One Step Hepatitis B Surface Antigen Test Device
particle coated with anti-HBsAg antibody. The mixture migrates (Serum/Plasma) will only indicate the presence of HBsAg in ORDERING INFORMATION
DIRECTIONS FOR USE
upward on the membrane chromatographically by capillary action Allow test device, serum or plasma specimen, and/or controls to the specimen and should not be used as the sole criteria for CATALOG NO. QUANTITY
to react with anti-HBsAg antibodies on the membrane and generate equilibrate to room temperature (15-30°C) prior to testing. the diagnosis of Hepatitis B viral infection.
a colored line. The presence of this colored line in the test region 3. As with all diagnostic tests, all results must be considered with 1160 001 25 test
1. Bring the pouch to room temperature before opening it.
indicates a positive result, while its absence indicates a negative other clinical information available to the physician. 1160 002 50 test
Remove the test device from the sealed pouch and use it as
result. To serve as a procedural control, a colored line will always soon as possible. Best results will be obtained if the assay is 4. The HBsAg One Step Hepatitis B Surface Antigen Test Device
appear in the control line region indicating that proper volume of performed within one hour. (Serum/Plasma) cannot detect less than 1 ng/mL of HBsAg in
specimen has been added and membrane wicking has occurred. 2. Place the test device on a clean and level surface. Hold the specimens. If the test result is negative and clinical symptoms
dropper vertically and transfer 3 full drops of serum or plasma persist, additional follow-up testing using other clinical methods
REAGENTS (approx. 100μl) to the specimen well (S) of the test device, and is suggested. A negative result at any time does not preclude
The test device contains anti-HBsAg particles and anti-HBsAg then start the timer. Avoid trapping air bubbles in the specimen the possibility of Hepatitis B infection.
coated on the membrane. well (S). See the illustration below.
3. Wait for the red line(s) to appear. The result should be read at EXPECTED VALUES
PRECAUTIONS 15 minutes. The HBsAg One Step Hepatitis B Surface Antigen Test Device
• For professional in vitro diagnostic use only. Do not use after Note: A low HBsAg concentration might result in a weak line (Serum/Plasma) has been compared with a leading commercial
expiration date. appearing in the test region (T) after an extended period of HBsAg EIA test. The correlation between these two systems is over
• Do not eat, drink or smoke in the area where the specimens time; therefore, do not interpret the result after 30 minutes. 98%.
and kits are handled.
• Handle all specimens as if they contain infectious agents. PERFORMANCE CHARACTERISTICS
Observe established precautions against microbiological Sensitivity
hazards throughout the procedure and follow the standard The HBsAg One Step Hepatitis B Surface Antigen Test Device
procedures for proper disposal of specimens. (Serum/Plasma) has been tested with a sensitivity panel ranging
• Wear protective clothing such as laboratory coats, disposable from 0 to 300 ng/mL. All 10 HBsAg subtypes produced positive
gloves and eye protection when specimens are assayed. results on the HBsAg One Step Hepatitis B Surface Antigen Test
• Humidity and temperature can adversely affect results. Device (Serum/Plasma). The test can detect 5ng/mL of HBsAg in
15 minutes, and 1 ng/mL of HBsAg in 30 minutes.
STORAGE AND STABILITY
The kit can be stored at room temperature or refrigerated (2-30°C).
The test device is stable through the expiration date printed on the
sealed pouch. The test device must remain in the sealed pouch
until use. DO NOT FREEZE. Do not use beyond the expiration date.
278 279
HBsAg/HCV Ab SYMBOLS IN PRODUCT LABELLING
11. Handle the negative and positive controls in the same manner
as patient specimens.
Rapid Test - Cassette EC REP Authorised Representative Temperature Limitation 12. The testing results should be read within 15 minutes after
(Serum/Plasma/Whole Blood) IVD For in-vitro diagnostic use Use by/Expiration Date a specimen is applied to the sample well or sample pad of
LOT Batch Code/Lot number CAUTION. Consult instructions the device. Reading the results after 15 minutes may give
erroneous results.
REF: 1162 001 25 test REF Catalogue Number for use
13. Do not perform the test in a room with strong air flow, i.e.
REF: 1162 002 50 test Consult instructions for use Manufactured by
electric fan or strong air-conditioning.
INTENDED USE REAGENT PREPARATION AND STORAGE

HBsAg antibody, and the C band is pre-coated with goat anti-mouse
The Spectrum HBsAg/HCV Ab Rapid Test is a lateral flow INSTRUCTIONS Step 5: Set up timer.
IgG antibody.
chromatographic immunoassay for the qualitative detection of All reagents are ready to use as supplied. Store unused test device
Hepatitis B surface antigen (HBsAg) and anti-Hepatitis C virus unopened at 2°C-30°C. Do not expose the kit over 30°C. Do not Step 6: Results can be read in 15 minutes. Positive results can
antibodies (IgG, IgM, IgA) in human serum, plasma and whole freeze the kit. The negative and positive controls should be store be visible in as short as 1 minute.
blood. It is intended to be used as a screening test and as an aid in at 2-8°C or the temperature indicated. If stored at 2°C-8°C, ensure
the diagnosis of infection with Hepatitis B virus (HBV) and Hepatitis that the test device is brought to room temperature before opening. Don’t read result after 15 minutes. To avoid confusion, discard
C virus (HCV). Any reactive specimen with the Spectrum HBsAg/ The test device is stable through the expiration date printed on the the test device after interpreting the result.
HCV Ab Rapid Test must be confirmed with alternative testing When an adequate volume of test specimen is dispensed into the
sealed pouch. QUALITY CONTROL
method(s) and clinical findings. sample well of the test cassette, the specimen migrates by capillary 1. Internal Control: This test contains a built-in control feature,
action across the cassette. The antibodies to HCV, if present in the SPECIMEN COLLECTION AND HANDLING the C line. The C line develops after adding the specimen and
Consider any materials of human origin as infectious and handle the sample diluent. If the C line does not develop, review the
SUMMARY AND EXPLANATION OF THE TEST specimen, will bind to the HCV Ag conjugates. The immunocomplex
is then captured on the membrane by the pre-coated non-conjugated them using standard biosafety procedures. whole procedure and repeat the test with a new device.
Hepatitis B virus (HBV) is the most common cause of persistent
HCV antigens on the HCV band forming a burgundy colored HCV 2. External Control: Good Laboratory Practice recommends
viremia and the most important cause of chronic liver disease and
band, indicating a HCV Ab positive or reactive test result. Plasma using external controls, positive and negative, to assure
hepatocellular carcinoma. It is estimated that there are 300 million
1. Collect blood specimen into a lavender, blue or green the proper performance of the assay, particularly under the
chronic carriers of HBV in the world. The carrier rates vary from as
HBsAg if present in the specimen will bind to the anti-HBsAg top collection tube (containing EDTA, citrate or heparin, following circumstances:
little as 0.3% (Western countries) to 20% (Asia, Africa)1.
conjugates. The immunocomplex is then captured on the membrane respectively, in Vacutainer® ) by veinpuncture. a. New operator uses the kit, prior to performing the testing
Hepatitis C, caused by Hepatitis C virus (HCV) infection, was by the pre-coated non-conjugated HBsAg antibody on the HBV 2. Separate the plasma by centrifugation. of specimens.
formerly described as the parenterally transmitted form of non-A, band forming a burgundy colored HBV band, indicating a HBsAg 3. Carefully withdraw the plasma into new pre-labeled tube. b. A new lot of test kits is used.
non-B hepatitis (NANBH). HCV can also be transmitted through positive test result. c. A new shipment of kits is used.
intravenous drug abuse, sexual, and household contact. About 3% Serum d. The temperature used during storage of the kits fall
of population is infected with HCV3, 15% of cases of infection are Absence of test bands suggests negative results. The test contains 1. Collect blood specimen into a red top collection tube (containing outside of 2-30°C.
acute and 80% of infections become chronic disease and are often an internal control (C band) which should exhibit a burgundy no anticoagulants in Vacutainer®) by veinpuncture. e. The temperature of the test area falls outside of 15-30°C.
associated with cirrhosis and hepatocellular carcinoma4. Cirrhosis colored band of the immunocomplex of goat anti-mouse IgG/anti- 2. Allow the blood to clot. f. To verify a higher than expected frequency of positive or
is more common in those co-infected with Hepatitis B virus. HBsAg conjugate regardless of the presence of colored test bands. 3. Separate the serum by centrifugation. negative results.
Otherwise, the test result is invalid and the specimen must be 4. Carefully withdraw the serum into a new pre-labeled tube. g. To investigate the cause of repeated invalid results.
Dual infection with HBV and HCV is not uncommon in geographic retested with another device.
areas where a high endemic level of both infections is reported Test specimens as soon as possible after collecting. Store
INTERPRETATION OF ASSAY RESULT
such as Southeast-Asia and the Mediterranean. In general, the REAGENTS AND MATERIALS PROVIDED specimens at 2°C to 8°C if not tested immediately. Store specimens
1. NEGATIVE RESULT: If only the C band is developed, the test
prevalence is around 10-20% in patients with chronic HBV infection, 1. Individually sealed foil pouches containing:: at 2°C to 8°C up to 5 days. The specimens should be frozen at
indicates that the levels of anti-HCV antibodies and HBsAg
and 2-10% of anti-HCV-positive patients have markers of HBV a. One cassette device -20°C for longer storage
in the specimen are undetectable. The result is negative or
infection. Co-infection of HBV and HCV was found to be high in b. One desiccant
Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen non-reactive.
HIV-infected people (66%), particularly in HIV infected drug users 2. Plastic droppers
(84%) 5, 6. specimens to room temperature slowly and mix gently. Specimens
3. Sample Diluent (1 vial, 5 mL)
containing visible particulate matter should be clarified by
4. One package insert (instruction for use)
Hepatitis B surface antigen (HBsAg) is the first marker to appear centrifugation before testing. Do not use samples demonstrating
in the blood in acute Hepatitis B, detected 1 week to 2 months gross lipemia, gross hemolysis or turbidity in order to avoid
after exposure and 2 weeks to 2 months before the onset of MATERIALS MAY BE REQUIRED AND NOT PROVIDED interference on result interpretation. 2. POSITIVE RESULT:
symptoms7. Three weeks after the onset of acute hepatitis almost 1. Positive Control I. If both C and HCV bands are developed, the test indicates
half of the patients will still be positive for HBsAg. In the chronic 2. Negative Control Blood the presence of antibodies to HCV in the specimen. The
carrier state, the HBsAg persists for long periods (6-12 months) Drops of whole blood can be obtained by either finger tip puncture result is HCV antibody positive.
with no seroconversion to the corresponding antibodies. Therefore, MATERIALS REQUIRED BUT NOT PROVIDED or veinpuncture. Do not use any hemolized blood for testing.
screening for HBsAg is highly desirable for all donors, pregnant 1. Clock or Timer
Whole blood specimens should be stored in refrigeration (2°C-8°C)
women and people in high-risk groups8. 2. Lancing device
if not tested immediately. The specimens must be tested within 24
Hepatitis C antibody (HCV Ab) tests such as ELISA and hours of collection.
WARNINGS AND PRECAUTIONS II. If both C and HBV bands are developed, the test indicates
chromatographic immunoassay are the most common tests for For in Vitro Diagnostic Use that the specimen contains HBsAg. The result is HBsAg
detection of HCV infection as detectable levels of HCV Ab are ASSAY PROCEDURE positive.
1. This package insert must be read completely before performing
generated about 6–8 weeks following infection 9-12. Step 1: Bring the specimen and test components to room
the test. Failure to follow the insert gives inaccurate test results.
temperature if refrigerated or frozen. Mix the specimen well prior to
The Spectrum HBsAg/HCV Ab Rapid Test is a point of care test 2. Do not open the sealed pouch unless ready to conduct the
assay once thawed.
which can detect both HBsAg and HCV antibodies in serum, plasma assay.
or whole blood in one device in 15 minutes by personnel with 3. Do not use expired devices.
Step 2: When ready to test, open the pouch at the notch and III. If C band, HCV band and HBV band are all developed,
minimal training and without laboratory equipment. 4. Bring all reagents to room temperature (15°C-30°C) before
remove device. Place the test device on a clean, flat surface. the test indicates the presence of antibodies to HCV and
use.
5. Do not use the components in any other type of test kit as a Step 3: Be sure to label the device with specimen’s ID number. HBsAg. The result is HCV antibody positive and HBsAg
TEST PRINCIPLE positive.
substitute for the components in this kit.
TThe Spectrum HBsAg/HCV Ab Rapid Test is a 3 line, lateral flow Step 4: Fill the plastic dropper with the specimen.
6. Do not use hemolized blood specimens for testing.
chromatographic immunoassay based on antibody sandwich assay
7. Wear protective clothing and disposable gloves while Holding the dropper vertically, dispense 1 drop of
(for HBsAg) and double antigen assay (for HCV Ab) test principles.
handling the kit reagents and clinical specimens. Wash hands specimen (about 40-50 µL for whole blood, 30-45 µL for
The test cassette consists of: 1) a burgundy colored conjugate
thoroughly after performing the test. serum/plasma) into the sample well making sure that
pad containing recombinant HCV core and non-structure antigens Samples with reactive results should be confirmed with alternative
8. Users of this test should follow the US CDC Universal there are no air bubbles.
conjugated with colloidal gold (HCV Ag conjugates) and mouse testing method(s) and clinical findings before a positive
Precautions for prevention of transmission of HIV, HBV and
anti-HBsAg antibody conjugated with colloidal gold (anti-HBsAg determination is made.
other blood-borne pathogens. Then add 1 drop (about 35-50 µL) of Sample Diluent.
conjugates), 2) a nitrocellulose membrane strip containing two
9. Do not smoke, drink or eat in areas where specimens or kit
test bands (HCV and HBV bands) and a control band (C band).
reagents are being handled.
The HCV band is pre-coated with recombinant HCV core and non-
10. Dispose of all specimens and materials used to perform the
structure antigens, the HBV band is pre-coated with non-conjugated
test as bio-hazardous waste.
280 281
3. INVALID: If no C line is developed, the assay is invalid Cross reaction with common microbe antigens: 5. If symptoms persist, while the result from Spectrum HBsAg/
regardless of color development on the T line as indicated The negative blood specimen was spiked with antigens from HCV Ab Rapid Test is non-reactive, it is recommended to re-
below. Repeat the assay with a new device. common microbes and then tested according to the standard sample the patient or test with an alternative test method.
procedure. The results showed that the Spectrum HBsAg/HCV Ab 6. Some specimens containing an unusually high titer of
Rapid Test had no cross-reaction with the following antigens at the heterophile antibodies or rheumatoid factor may affect
concentration tested. expected results.
Concentration HCV Ab HBsAg findings.
Antigen (Ag)
spiked Reactivity Reactivity
PERFORMANCE CHARACTERISTICS REFERENCES
1. Clinical Performance HIV P24 Ag 1.0 mg/mL Negative Negative 1. Maheshwari, A; Ray S; Thuluvath PJ (2008-07-26). “Acute
A total of 1050 samples from susceptible subjects were tested with hepatitis C”. Lancet 372 (9635): 321–32.
the Spectrum HBsAg/HCV Ab Rapid Test and by a commercial 2. Nelson, PK; Mathers BM, et,al (2011-08-13). “Global
Dengue virus NS1 epidemiology of hepatitis B and hepatitis C in people who
HCV Ab ELISA kit. Comparison for all subjects is shown in the 1.0 mg/mL Negative Negative
Ag (I, II, III,IV) inject drugs: results of systematic reviews”. Lancet 378 (9791):
following table
571–83
Chikungunya Ag 1.0 mg/ml Negative Negative 3. Liu Z, Hou J. Hepatitis B Virus (HBV) and Hepatitis C Virus
Spectrum HBsAg/HCV Ab (HCV) Dual Infection. Int J Med Sci 2006; 3(2):57-62
Rapid Test 4. Magnius LO, Espmark A. A new antigen complex co-occurring
3. Interference with Australia antigen. Acta Pathol Microbiol Scand [B]
HCV Ab ELISA Positive Negative Total Common substances (such as pain and fever medication, Microbiol Immunol. 1972;80(2):335-7
blood components) may affect the performance of the Spectrum 5. Kao JH. Diagnosis of hepatitis B virus infection through
Positive 312 4 316 serological and virological markers. Expert Rev Gastroenterol
HBsAg/HCV Ab Rapid Test. This was studied by spiking of these
substances to the three levels of the HCV Ab and HBsAg. The Hepatol. 2008;2(4):553-562
Negative 3 731 734
results are presented on the following table and demonstrate 6. Esteban JI, Gonzalez A, Hernandez JM, et al. Evaluation
Total 315 735 1050 that the substances studied did not affect the performance of the of antibodies to hepatitis C virus in a study of transfusion-
Spectrum HBsAg/HCV Ab Rapid Test. associated hepatitis. N Engl J Med 1990. 323:1
HCV Ab Relative Sensitivity: 98.7%, Relative Specificity: 99.6%, 7. Caruntu FA, Benea L. Acute hepatitis C virus infection:
Overall Agreement: 99.3% Note: -: Negative; +: Weak Positive; +++: Strong Positive Diagnosis, pathogenesis, treatment. J. Gastrointestin Liver
Dis. 2006; 15(3):249-256.
A total of 1056 samples from susceptible subjects were tested with Potential HCV Ab Reactivity HBsAg Reactivity
the Spectrum HBsAg/HCV Ab Rapid Test and by a commercial interfering
HBsAg ELISA kit. Comparison for all subjects is shown in the substances Weak Strong Weak Strong
Negative Negative
following table. spiked Positive Positive Positive Positive
Control - + +++ - + +++

Spectrum HBsAg/HCV Ab Rapid
Test Bilirubin
- + +++ - + +++
20 mg/dL
HBsAg ELISA Positive Negative Total
Creatinine
- + +++ - + +++
Positive 322 0 322 442 mmol/L
Glucose
Negative 2 732 734 - + +++ - + +++
55 mmol/L
Albumin
Total 324 732 1056 - + +++ - + +++
60 g/L
Salicylic
HBsAg relative Sensitivity: 100%, Relative Specificity: 99.7%,
acid 4.34 - + +++ - + +++
Overall Agreement: 99.8%
mmol/L
2. Cross-Reactivity Heparin
- + +++ - + +++
Cross-reactivity with specimens from other infectious diseases: 3,000 U/L
Specimens from other common infectious diseases were collected EDTA
- + +++ - + +++
and tested with the Spectrum HBsAg/HCV Ab Rapid Test for a cross 3.4 mmol/L
reaction study. The result is shown in the table below: Human IgG
- + +++ - + +++
150 mg/dL
Sample HCV Ab HBsAg
Specimens
size Reactivity Reactivity
LIMITATIONS OF TEST
HAV positive serum 10 Negative Negative 1. The Assay Procedure and the Interpretation of Assay Result
must be followed closely when testing for the presence of anti-
HIV positive serum 10 Negative Negative HCV antibodies and/or HBsAg in serum, plasma and whole
blood from individual subjects. Failure to follow the procedure
HBsAg positive serum 10 Negative Positive may give inaccurate results.
2. The Spectrum HBsAg/HCV Ab Rapid Test is limited to the
HCV Ab positive serum 10 Positive Negative qualitative detection of anti HCV antibodies and/or HBsAg in
human serum, plasma and whole blood. The intensity of the
Syphilis positive serum 10 Negative Negative
test bands does not have linear correlation with anti-HCV
TB positive serum 10 Negative Negative antibodies titer or HBsAg titer in the specimen.
3. A nonreactive test result does not preclude the possibility of
H. pylori positive serum 10 Negative Negative exposure to or infection with HCV and/or HBV.
4. A nonreactive result can occur if the quantity of anti-HCV
ANA serum 8 Negative Negative antibody and/or HBsAg present in the specimen is below the
detection limits of the assay, or anti-HCV antibody and/or the
RF (≤2,500 IU/ml) serum 3 Negative Negative HBsAg that are detected are not present during the stage of
disease in which a sample is collected.
HAMA specimens 19 Negative Negative
282 283
HCG date printed on the sealed pouch. The test must remain in the
sealed pouch until use. DO NOT FREEZE. Do not use beyond the
(C). No apparent colored line appears in the test line region (T).
INVALID: Control line fails to appear. Insufficient specimen volume
Device (Urine/Serum) and another commercially available
urine/serum membrane hCG test. The urine study included 159
One Step Pregnancy Test Device (Urine/Serum) expiration date. or incorrect procedural techniques are the most likely reasons for specimens, and both assays identified 88 negative and 71 positive
control line failure. Review the procedure and repeat the test with results. The serum study included 73 specimens and both assays
REF: 1176 001 25 test SPECIMEN COLLECTION AND PREPARATION a new test. If the problem persists, discontinue using the test kit identified 51 negative, 21 positive and 1 invalid results. The
REF: 1176 002 50 test Urine Assay immediately and contact your local distributor. results demonstrated a >99% overall accuracy of the hCG One
A urine specimen must be collected in a clean and dry container. A Step Pregnancy Test Device (Urine/Serum) when compared to the
A rapid, one step test for the qualitative detection of human chorionic first morning urine specimen is preferred since it generally contains QUALITY CONTROL other urine/serum membrane hCG test.
gonadotropin (HCG) in urine or serum. the highest concentration of hCG; however, urine specimens A procedural control is included in the test. A colored line appearing
collected at any time of the day may be used. Urine specimens in the control line region (C) is considered an internal procedural hCG Reference Method (Urine)
INTENDED USE exhibiting visible precipitates should be centrifuged, filtered, or control. It confirms sufficient specimen volume and correct Method Other hCG Rapid Test
The hCG One Step Pregnancy Test Device (Urine/Serum) is a Total
allowed to settle to obtain a clear specimen for testing. procedural technique. A clear background is an internal negative
rapid chromatographic immunoassay for the qualitative detection of Results Positive Negative Results
procedural control. If a background color appears in the result hCG
human chorionic gonadotropin in urine or serum to aid in the early Serum Assay window and interferes with the ability to read the test result, the Test Positive 71 0 71
detection of pregnancy. Blood should be collected aseptically into a clean tube without result may be invalid. Device
Negative 0 88 88
anticoagulants. Separate the serum from blood as soon as possible It is recommended that a positive hCG control (containing 10-250
SUMMARY to avoid hemolysis. Use clear non-hemolyzed specimens when mIU/mL hCG) and a negative hCG control (containing “0” mIU/ Total Results 71 88 159
Human chorionic gonadotropin (hCG) is a glycoprotein hormone possible. mL hCG) be evaluated to verify proper test performance when a
produced by the developing placenta shortly after fertilization. In new shipment of tests are received. Sensitivity: 100% (96%-100%)* Specificity: 100% (95%-100%)*
normal pregnancy, hCG can be detected in both urine and Specimen Storage Accuracy: 100% (98%-100%)*
serum as early as 7 to 10 days after conception. hCG levels Urine or serum specimens may be stored at 2-8 OC for up to 48 LIMITATION * 95% Confidence Intervals
continue to rise very rapidly, frequently exceeding 100 mIU/ hours prior to testing. For prolonged storage, specimens may be 1. The hCG One Step Ultra Pregnancy Test Device (Urine/
mL by the first missed menstrual period and peaking in the100, frozen and stored below -20 OC. Frozen specimens should be Serum) is a preliminary qualitative test, therefore, neither the hCG Reference Method (Serum)
000-200,000 mIU/mL range about 10-12 weeks into pregnancy. thawed and mixed before testing. quantitative value nor the rate of increase in hCG can be
The appearance of hCG in both the urine and serum soon after Method Other hCG Rapid Test Total
determined by this test.
conception, and its subsequent rapid rise in concentration during MATERIALS Results Positive Negative Results
2. Very dilute urine specimens, as indicated by a low specific hCG
early gestational growth, make it an excellent marker for the early Materials Provided gravity, may not contain representative levels of hCG. If Test Positive 21 0 21
detection of pregnancy. • Test devices pregnancy is still suspected, a first morning urine specimen Device
The hCG One Step Pregnancy Test Device (Urine/Serum) is a • Droppers should be collected 48 hours later and tested. Negative 0 51 51
rapid test that qualitatively detects the presence of hCG in urine or • Package inserts 3. Very low levels of hCG (less than 50 mIU/mL) are present Total Results 21 51 72
serum specimens at the sensitivity of 10 mIU/mL. The test utilizes a in urine and serum specimen shortly after implantation.
combination of monoclonal and polyclonal antibodies to selectively Materials Required But Not Provided
However, because a significant number of first trimester Sensitivity: 100% (86%-100%)* Specificity: 100% (93%-100%)*
detect elevated levels of hCG in urine or serum. At the level of • Specimen collection containers
pregnancies terminate for natural reasons, a test result that is Accuracy: 100% (95%-100%)*
claimed sensitivity, the hCG One Step Ultra Pregnancy Test Device • Timer
weakly positive should be confirmed by retesting with a first * 95% Confidence Intervals
(Urine/Serum) shows no cross-reactivity interference from the morning urine or serum specimen collected 48 hours later.
structurally related glycoprotein hormones hFSH, hLH and hTSH at DIRECTIONS FOR USE 4. This test may produce false positive results. A number of
high physiological levels. Allow test, urine or serum specimen and/or controls to equili- Sensitivity and Specificity
conditions other than pregnancy, including trophoblastic The hCG One Step Pregnancy Test Device (Urine/Serum) detects
brate to room temperature (15-30 OC) prior to testing. disease and certain non-trophoblastic neoplasms including
1. Bring the pouch to room temperature before opening it. hCG at a concentration of 25 mIU/mL or greater. The test has been
PRINCIPLE testicular tumors, prostate cancer, breast cancer, and lung standardized to the W.H.O. International Standard. The addition
The hCG One Step Ultra Pregnancy Test Device (Urine/Serum) is Remove the test device from the sealed pouch and use it as cancer, cause elevated levels of hCG. Therefore, the presence
soon as possible. of LH (300 mIU/mL), FSH (1,000 mIU/mL), and TSH (1,000 µIU/
a rapid chromatographic immunoassay for the qualitative detection of hCG in urine or serum specimens should not be used to
2. Place the test device on a clean and level surface. Hold the mL) to negative (0 mIU/mL hCG) and positive (25 mIU/mL hCG)
of human chorionic gonadotropin in urine or serum to aid in the diagnose pregnancy unless these conditions have been ruled
dropper vertically and transfer 3 full drops of urine or serum specimens showed no cross- reactivity.
early detection of pregnancy. The test uses two lines to indicate out.
results. The test line utilizes a combination of antibodies including (approx. 100 µL) to the specimen well (S) of the test device, 5. This test may produce false negative results. False negative
a monoclonal HCG antibody to selectively detect elevated levels and then start the timer. Avoid trapping air bubbles in the results may occur when the levels of hCG are below the
of hCG. The control line is composed of goat polyclonal antibodies specimen well (S). See the illustration below. The following potentially interfering substances were added to
sensitivity level of the test. When pregnancy is still suspected,
and colloidal gold particles. The assay is conducted by adding a 3. Wait for the colored line(s) to appear. Read the result at 3 hCG negative and positive specimens.
a first morning urine or serum specimen should be collected 48
urine or serum specimen to the specimen well of the test device and minutes when testing a urine specimen, or at 5 minutes hours later and tested. In case pregnancy is suspected and the Acetaminophen 20 mg/dL Caffeine 20 mg/dL
observing the formation of colored lines. The specimen migrates when testing a serum specimen. test continues to produce negative results, see a physician for Acetylsalicylic Acid 20 mg/dL Gentisic Acid 20 mg/dL
via capillary action along the membrane to react with the colored NOTE: A low hCG concentration might result in a weak line further diagnosis. Ascorbic Acid 20 mg/dL Glucose 2 g/dL
conjugate. appearing in the test region (T) after an extended period of 6. As with any assay employing mouse antibodies, the possibility
Positive specimens react with the specific antibody-hCG-colored time; therefore, do not interpret the result after 10 minutes. exists for interference by human anti-mouse antibodies Atropine 20 mg/dL Hemoglobin 1 mg/dL
conjugate to form a colored line at the test line region of the (HAMA) in the specimen. Specimens from patients who have Bilirubin (serum) 40 mg/dL Bilirubin (urine) 2 mg/dL
membrane. Absence of this colored line suggests a negative result. received preparations of monoclonal antibodies for diagnosis
To serve as a procedural control, a colored line will always appear Triglycerides (serum) 1,200 mg/dL
or therapy may contain HAMA. Such specimens may cause
in the control line region indicating that proper volume of specimen false positive or false negative results. None of the substances at the concentration tested interfered in the
has been added and membrane wicking has occurred. 7. This test provides a presumptive diagnosis for pregnancy. A assay.
confirmed pregnancy diagnosis should only be made by a
REAGENTS physician after all clinical and laboratory findings have been BIBLIOGRAPHY
The test contains anti-hCG particles and anti-hCG coated on the evaluated. 1. Batzer FR. Hormonal evaluation of early pregnancy, Fertil.
membrane. Steril. 1980; 34(1): 1-13
EXPECTED VALUES 2. Catt KJ, ML Dufau, JL Vaitukaitis Appearance of hCG in
Negative results are expected in healthy non-Pregnant women and pregnancy plasma following the initiation of implantation of the
PRECAUTIONS
healthy men. blastocyte, J. Clin. Endocrinol. Metab. 1975; 40(3): 537-540
• For professional in vitro diagnostic use only. Do not use after
Healthy Pregnant women have hCG present in their urine and serum 3. Braunstein GD, J Rasor, H. Danzer, D Adler, ME Wade
the expiration date.
specimens. The amount of hCG will vary greatly with gestational Serum human chorionic gonadotropin levels throughout
• The test should remain in the sealed pouch until use. INTERPRETATION OF RESULTS
age and between individuals. normal pregnancy, Am. J. Obstet. Gynecol.
• Do not use test if the package is damaged. (Please refer to the illustration above)
• All specimens should be considered potentially hazardous and The hCG one step pregnancy test device (urine/serum) has a
POSITIVE: *Two distinct colored lines appear. One line should be
handled in the same manner as an infectious agent. sensitivity of 25 mIU/mL, and is capable of detecting pregnancy as ORDERING INFORMATION
in the control line region (C) and another line should be in the test
• The used test should be discarded according to local early as 1 day after the first missed menses.
line region (T). CATALOG NO. QUANTITY
regulations. *NOTE: The intensity of the color in the test line region (T) may vary
depending on the concentration of hCG present in the specimen. PERFORMANCE CHARACTERISTICS 1176 001 25 test
STORAGE AND STABILITY Therefore, any shade of color in the test line region (T) should be 1176 002 50 test
Accuracy
Store as packaged in the sealed pouch at room temperature or considered positive. A multi-center clinical evaluation was conducted comparing the
refrigerated (2-30OC). The test is stable through the expiration NEGATIVE: : One colored line appears in the control line region results obtained using the hCG One Step Pregnancy Test
284 285
HCV Ab SYMBOLS IN PRODUCT LABELLING
EXPECTED VALUES
The HCV One Step Test Device (Serum/Plasma) has been
Hepatitis C Virus Ab Test Device EC REP Authorised Representative Temperature Limitation compared with a leading commercial HCV EIA test. The correlation
(Serum/Plasma) IVD For in-vitro diagnostic use Use by/Expiration Date between these two systems is 98%.
LOT Batch Code/Lot number CAUTION. Consult
REF: 1164 001 25 test Catalogue Number instructions for use PERFORMANCE CHARACTERISTICS
REF
REF: 1164 002 50 test
Consult instructions for use Manufactured by Sensitivity
A rapid, one step test for the qualitative detection of antibodies to The HCV One Step Test Device (Serum/Plasma) has passed a
Hepatitis C Virus in serum or plasma. seroconversion panel and compared with a leading commercial
For professional in vitro diagnostic use only. HCV EIA test using clinical specimens.
Specificity
INTENDED USE SPECIMEN COLLECTION AND PREPARATION The recombinant antigen used for the HCV One Step Test
The HCV One Step Test Device (Serum/Plasma) is a rapid • The HCV One Step Test Device (Serum/Plasma) can be
Device (Serum/Plasma) is encoded by genes for both structural
chromatographic immunoassay for the qualitative detection of performed using either serum or plasma.
(nucleocapsid) and non-structural proteins. The HCV One Step Test
antibody to Hepatitis C Virus in serum or plasma. • Separate the serum or plasma from blood as soon as possible
Device (Serum/Plasma) is highly specific for antibodies to Hepatitis
to avoid hemolysis. Only clear, non-hemolyzed specimens can
C Virus compared with a leading commercial HCV EIA test.
SUMMARY be used.
Hepatitis C Virus (HCV) is a small, enveloped, positive-sense, • Testing should be performed immediately after the specimens
HBsAg Reference Method
single-stranded RNA Virus. HCV is now known to be the major have been collected. Do not leave the specimens at room
temperature for prolonged periods. Specimens may be stored Method EIA Total
cause of parenterally transmitted non-A, non-B hepatitis. Antibody
to HCV is found in over 80% of patients with well-documented at 2-8°C for up to 3 days. For long term storage, specimens Results Positive Negative Results
should be kept below -20°C. HCV
non-A, non-B hepatitis. Conventional methods fail to isolate the
• Bring specimens to room temperature prior to testing. Frozen Test Positive 92 20 112
virus in cell culture or visualize it by electron microscope. Cloning
specimens must be completely thawed and mixed well prior Device
the viral genome has made it possible to develop serologic assays Negative 3 1888 1891
that use recombinant antigens.1,2 Compared to the first generation to testing. Specimens should not be frozen and thawed INTERPRETATION OF RESULTS
repeatedly. (Please refer to the illustration above) Total Results 95 1908 2003
HCV EIAs using single
• If specimens are to be shipped, they should be packed in POSITIVE: * Two distinct red lines appear. One line should be in the
recombinant antigen, multiple antigens using recombinant protein Relative sensitivity: 96.8%
compliance with federal regulations for transportation of control region (C) and another line should be in the test region (T).
and/or synthetic peptides have been added in new serologic tests Relative specificity: 99.0%
to avoid nonspecific cross-reactivity and to increase the sensitivity etiologic agents. *NOTE: The intensity of the red color in the test line region (T) may
vary depending on the concentration of HCV antibodies present in Accuracy: 98.9%
of the HCV antibody tests.3,4
The HCV One Step Test Device (Serum/Plasma) is a rapid test to the specimen. Therefore, any shade of red in the test region should Precision
MATERIALS be considered positive.
qualitatively detect the presence of antibody to HCV in a serum Materials Provided Intra-Assay
or plasma specimen. The test utilizes a combination of protein A NEGATIVE: One red line appears in the control region (C). No Within-run precision has been determined by using 15 replicates
• Test devices
coated particles and recombinant HCV proteins to selectively detect apparent red or pink line appears in the test region (T). of three specimens: a negative, a low positive and a high positive.
• Disposable specimen droppers
antibody to HCV in serum or plasma. The recombinant HCV proteins INVALID: Control line fails to appear. Insufficient specimen volume The negative, low positive and high positive values were correctly
• Buffer
used in the test kit are encoded by the genes for both structural or incorrect procedural techniques are the most likely reasons for identified 98% of the time.
• Package insert
(nucleocapsid) and non-structural proteins. control line failure. Review the procedure and repeat the test with a
Materials Required But Not Provided new test device. If the problem persists, discontinue using the test Inter-Assay
• Specimen collection container kit immediately and contact your local distributor. Between-run precision has been determined by 15 independent
PRINCIPLE assays on the same three specimens: a negative, a low positive
• Pipette and disposable tips (optional)
The HCV One Step Test Device (Serum/Plasma) is a qualitative, and a high positive. Three different lots of the HCV One Step Test
• Centrifuge (for plasma only) QUALITY CONTROL
membrane based immunoassay for the detection of antibody Device (Serum/Plasma) have been tested over a 3-month period
• Timer
to HCV in serum or plasma. The membrane is coated with Internal Quality Control using negative, low
recombinant HCV antigen on the test line region of the device. A procedural control is included in the test. A colored line appearing positive and high positive specimens. The specimens were correctly
During testing, the serum or plasma specimen reacts with the DIRECTIONS FOR USE
Allow test device, specimen, buffer and/or controls to equilibrate to in the control line region (C) is considered an internal procedural identified 98% of the time.
Protein A coated particles. The mixture migrates upward on the control. It confirms sufficient specimen volume, adequate membrane
membrane chromatographically by capillary action to react with room temperature (15-30°C) prior to testing.
1. Remove the test device from the foil pouch and use it as wicking and correct procedural technique. BIBLIOGRAPHY
recombinant HCV antigen on the membrane and generate a colored
soon as possible. Best results will be obtained if the assay is External Quality Control 1. Kuo, G., Q.L. Choo, H.J. Alter, and M. Houghton. An assay
line. Presence of this colored line indicates a positive result, while
performed within one hour. It is recommended that a positive and negative external control be for circulating antibodies to a major etiologic Virus of human
its absence indicates a negative result. To serve as a procedural
2. Place the test device on a clean and level surface. Transfer the run every 20 tests, and as deemed necessary by internal laboratory non-A, non-B hepatitis. Science 1989; 244:362
control, a colored line will always appear at the control line region
indicating that proper volume of specimen has been added and specimen by a pipette or a dropper: procedures. Some commercial controls may contain interfering
• To use a Pipette: Transfer 5 μL of serum or plasma to preservatives; therefore, interpretation of results with external 2. Choo, Q.L., G. Kuo, A.J. Weiner, L.R. Overby, D.W. Bradley,
membrane wicking has occurred.
the specimen well (S) of the test device, then add 2 full quality controls should be done with caution. and M. Houghton. Isolation of a cDNA clone derived from a
drops of buffer (approximately 80 μL) and start the timer. blood-borne non-A, non-B viral hepatitis genome. Science
REAGENTS Procedure for External Quality Control Testing 1989; 244:359
Avoid trapping air bubbles in the specimen well (S). See
The test device contains protein A coated particles and HCV antigen 1. Using a disposable dropper or pipette, transfer 5 μL to the
illustration 1 below.
coated on the membrane. Specimen well (S) as in procedure Step 2.
• To use a Disposable Specimen Dropper: Hold the 3. Wilber, J.C. Development and use of laboratory tests for
dropper vertically, draw the specimen up to the Fill Line 2. Continue with Step 3 of Directions For Use. If the controls hepatitis C infection: a review. J.Clin. Immunoassay 1993;
PRECAUTIONS as shown in illustration 2 below (approximately 5 μL). do not yield the expected results, do not use the test results. 16:204
• For professional in vitro diagnostic use only. Do not use after Transfer the specimen to the specimen well (S) of the Repeat the test or contact your distributor.
expiration date. test device, then add 2 full drops of buffer (approximately 4. van der Poel, C. L., H.T.M. Cuypers, H.W. Reesink, and
• Do not eat, drink or smoke in the area where the specimens 80 μL) and start the timer. Avoid trapping air bubbles in LIMITATION P.N.Lelie. Confirmation of hepatitis C Virus infection by new
and kits are handled. the specimen well (S). 1. The HCV One Step Test Device (Serum/Plasma) is for in vitro four-antigen recombinant immunoblot assay. Lancet 1991;
• Handle all specimens as if they contain infectious agents. 3. Wait for the red line(s) to appear. The result should be read at diagnostic use only. This test should be used for the detection 337:317
Observe established precautions against microbiological 10 minutes. Do not interpret the result after 20 minutes. of antibodies to HCV in serum or plasma specimen.
hazards throughout the procedure and follow the standard 2. The HCV One Step Test Device (Serum/Plasma) will only
procedures for proper disposal of specimens. indicate the presence of antibodies to HCV in the specimen
• Wear protective clothing such as laboratory coats, disposable and should not be used as the sole criteria for the diagnosis of
gloves and eye protection when specimens are assayed. Hepatitis C viral infection.
• Humidity and temperature can adversely affect results. ORDERING INFORMATION
3. As with all diagnostic tests, all results must be considered with
other clinical information available to the physician. CATALOG NO. QUANTITY
STORAGE AND STABILITY 4. If the test result is negative and clinical symptoms persist, 1164 001 25 test
The kit can be stored at room temperature or refrigerated (2-30°C). additional follow-up testing using other clinical methods is 1164 002 50 test
The test device is stable through the expiration date printed on the recommended. A negative result at any time does not preclude
sealed pouch. The test device must remain in the sealed pouch the possibility of Hepatitis C Virus infection.
until use. DO NOT FREEZE. Do not use beyond the expiration date.
286 287
HIV Ab 1/2/O
Tri-line Human Immunodeficiency Virus Rapid Test Device
STORAGE AND STABILITY
Store as packaged in the sealed pouch either at room temperature or
start the timer. See illustration below. For Fingerstick Whole
Blood specimens:
EXPECTED VALUES
The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid Test
refrigerated (2-30°C). The test device is stable through the expiration • To use a capillary tube: Fill the capillary tube and transfer Device (Whole Blood/Serum/Plasma) has been compared with
(Whole Blood/Serum/Plasma) date printed on the sealed pouch. The test device must remain in the approximately 50 μL of fingerstick whole blood leading commercial HIV ELISA tests and/or Western Blot. The
sealed pouch until use. DO NOT FREEZE. Do not use beyond the specimen to the specimen well (S) of the test device, correlation between these two systems is 99.8%.
REF: 1168 001 25 test expiration date. then add 2 drops of buffer (approximately 80 μL) and
REF: 1168 002 50 test start the timer. See illustration below. PERFORMANCE CHARACTERISTICS
SPECIMEN COLLECTION AND PREPARATION • To use hanging drops: Allow 2 hanging drops of Clinical Sensitivity, Specificity and Accuracy
A rapid test for the qualitative detection of antibodies to Human • The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid fingerstick whole blood specimen (approximately 50 The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid
Immunodeficiency Virus (HIV) type 1, type 2, and Subtype O in Test Device (Whole Blood/Serum/Plasma) can be performed μL) to fall into the center of the specimen well (S) on the Test Device (Whole Blood/Serum/Plasma) was evaluated in a
whole blood, serum or plasma. using whole blood (from venipuncture or fingerstick), serum or test device, then add 2 drops of buffer (approximately multi-center field study, a blood donation center as well as an
For professional in vitro diagnostic use only. plasma. 80 μL) and start the timer. See illustration below. in-house clinical study. The multi-center study included 1,640
• To collect Fingerstick Whole Blood specimens: 3. Wait for the colored line(s) to appear. Read results at 10 specimens from different countries. There were 1,000 specimens
INTENDED USE • Wash the patient’s hand with soap and warm water or minutes. Do not read results after 20 minutes. from the blood donation center, and the in-house clinical study
The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid Test clean with an alcohol swab. Allow to dry. included 687 specimens and an HIV Performance Panel that was
Device (Whole Blood/Serum/Plasma) is a rapid chromatographic • Massage the hand without touching the puncture site by purchased from a commercial source. The HIV 1/2/O Tri-line Human
immunoassay for the qualitative detection of antibodies to HIV-1, rubbing down the hand towards the fingertip of the middle Immunodeficiency Virus Rapid Test Device (Whole Blood/Serum/
HIV-2, and Subtype O in whole blood, serum or plasma to aid in the or ring finger. Plasma) was compared to leading commercial ELISA HIV tests
diagnosis of HIV infection. • Puncture the skin with a new sterile lancet for each and/or Western Blot. Of the 3,327 total specimens, 872 were found
person. Wipe away the first sign of blood. positive and 2,455 specimens were found negative by ELISA and/or
SUMMARY • Gently rub the hand from wrist to palm to finger to form a Western Blot. The HIV 1/2/O Tri-line Human Immunodeficiency Virus
HIV is the etiologic agent of Acquired Immune Deficiency Syndrome rounded drop of blood over the puncture site. Rapid Test Device (Whole Blood/Serum/Plasma) showed 99.9%
(AIDS). The virion is surrounded by a lipid envelope that is derived • Add the Fingerstick Whole Blood specimen to the test relative sensitivity, and 99.8% relative specificity compared to ELISA
from the host cell membrane. Several viral glycoproteins are on the device by using a capillary tube: and/or Western Blot.
envelope. Each virus contains two copies of positive-sense genomic • Touch the end of the capillary tube to the blood until
RNAs. HIV-1 has been isolated from patients with AIDS and AIDS- filled to approximately 50 μL. Avoid air bubbles. HIV 1/2/O Tri-line Rapid Test Device vs. ELISA and/or Western Blot
related complex, and from healthy people with high potential risk • Place the bulb onto the top end of the capillary
for developing AIDS.1 HIV-1 consists of Subtype M and Subtype O. tube, then squeeze the bulb to dispense the whole ELISA and/or Western
Method Total
Highly blood into the specimen well (S) of the test device. Blot
divergent strains of HIV-1 were first recognized in 1990 and grouped • Add the Fingerstick Whole Blood specimen to the test Results
provisionally as Subtype O as this variation has similar glycoprotein HIV 1/2/O Results Positive Negative
device by using hanging drops: Tri-line
markers to HIV-1 but a slight variation to the protein marker. • Position the patient’s finger so that the drop of Positive 871 6 877
Rapid
Although rarely compared to HIV-1 and HIV-2, infections caused by blood is just above the specimen well (S) of the Test
Subtype O have so far been identified in Africa (Cameroon), France test device. Negative 1 2449 2450
Device
and Germany. HIV-2 has been isolated from West African AIDS • Allow 2 hanging drops of fingerstick whole blood
patients and from seropositive asymptomatic individuals.2 HIV-1, to fall into the center of specimen well (S) on the Total Results 872 2455 3327
HIV-2, and Subtype O all elicit immune responses.3 Detection of test device, or move the patient’s finger so that the
HIV antibodies in serum, plasma or whole blood is the most efficient hanging drop touches the center of the specimen Relative Sensitivity: 99.9% (99.4-100 %)* Relative Specificity: 99.8%
and common way to determine whether an individual has been
INTERPRETATION OF RESULTS
well (S). Avoid touching the finger directly to the (Please refer to the illustration above) (99.5-99.9%)*
exposed to HIV and to screen blood and blood products for HIV.4 specimen well (S). Relative Accuracy: 99.8% (99.6-99.9%)* * 95% Confidence Interval
POSITIVE:* Two or three distinct colored lines appear. One line
Despite the differences in their biological characters, serological • Separate serum or plasma from blood as soon as possible to should always appear in the control line region (C), and another
activities and genome sequences, HIV-1, HIV-2, and Subtype O avoid hemolysis. Use only clear, nonhemolyzed specimens. Precision
one or two apparent colored line(s) should appear in the test line
show strong antigenic crossreactivity. 5,6 Most HIV-2 positive sera • Testing should be performed immediately after specimen Intra-Assay
region(s) (T1 and/or T2).
can be identified by using HIV-1 based serological tests. The HIV collection. Do not leave the specimens at room temperature Within-run precision has been determined by using 10 replicates of
*NOTE: The intensity of the color in the test line region (T1 and T2)
1/2/O Tri-line Human Immunodeficiency Virus Rapid Test Device for prolonged periods. Serum and plasma specimens may four specimens: a negative, a low positive, medium positive and a
will vary depending on the concentration of HIV antibodies present
(Whole Blood/Serum/Plasma) is a rapid test to qualitatively detect be stored at 2-8°C for up to 3 days. For long term storage, high positive. The negative, low positive, medium positive and high
in the specimen. Therefore, any shade of color in the test line region
the presence of antibodies to HIV-1, HIV-2, and/or Subtype O in specimens should be kept below -20°C. Whole blood collected positive values were correctly identified >99% of the time.
(T1 and/or T2) should be considered positive.
whole blood, serum or plasma specimen. by venipuncture should be stored at 2-8°C if the test is to be NEGATIVE: One colored line appears in the control region (C).
run within 2 days of collection. Do not freeze whole blood No apparent colored lines appear in the test line regions (T1 and T2). Inter-Assay
PRINCIPLE Between-run precision has been determined by 10 independent
The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid Test specimens. Whole blood collected by fingerstick should be INVALID: Control line fails to appear. Insufficient specimen volume
tested immediately. or incorrect procedural techniques are the most likely reasons for assays on the same four specimens: a negative, a low positive,
Device (Whole Blood/Serum/Plasma) is a qualitative, membrane
• Bring specimens to room temperature prior to testing. Frozen control line failure. Review the procedure and repeat the test with a medium positive and a high positive. Three different lots of the HIV
based immunoassay for the detection of antibodies to HIV-1, HIV-2,
specimens must be completely thawed and mixed well prior to new test device. If the problem persists, discontinue using the test kit 1/2/O Tri-line Human Immunodeficiency Virus Rapid Test Device
and Subtype O in whole blood, serum or plasma. The membrane is
testing. Specimens should not be frozen and thawed repeatedly. immediately and contact your local distributor. (Whole Blood/Serum/Plasma) have been tested using negative,
pre-coated with recombinant HIV antigens in the test line regions,
• If specimens are to be shipped, they should be packed in low positive, medium positive and high positive specimens. The
T1 and T2. The T1 test line is pre-coated with HIV-1 and Subtype O
antigen and the T2 test line is pre-coated with HIV-2 antigen. During compliance with local regulations covering the transportation of QUALITY CONTROL specimens
etiologic agents. A procedural control is included in the test. A colored line appearing were correctly identified >99% of the time.
testing, the whole blood, serum or plasma specimen reacts with the
in the control line region (C) is considered an internal procedural
mixture of HIV-1 envelope and core antigens and HIV-2 envelope MATERIALS BIBLIOGRAPHY
control. It confirms sufficient specimen volume, adequate membrane
antigen that are coated on colored particles in the test strip. The Materials Provided 1. Janssen RS, Satten,GA, Stramer SL, Rawal BD, O’Brien
wicking and correct procedural technique.
mixture then migrates upward on the membrane chromatographically • Test devices TR, Weiblen BJ, Hecht FM, Jack N, Cleghorn FR, Kahn JO,
Control standards are not supplied with this kit; however, it is
by capillary action and reacts with recombinant HIV antigen on the • Droppers Chesney MA and Busch MP. New testing strategy to detect
recommended that positive and negative controls be tested as a
membrane in the test line region. If the specimen contains antibodies • Buffer early HIV-1 infection for use in incidence estimates and for
good laboratory practice to confirm the test procedure and to verify
to HIV-1 and/or Subtype O, or HIV-2, one colored line will appear • Package insert clinical and prevention purposes. JAMA (1998) 280(1):42-48.
proper test performance.
in the test line region; if the specimen contains antibodies to HIV-1 2. Chang SY, Bowman BH, Weiss JB, Garcia RE and White TJ.
and/or Subtype O, and HIV-2, two colored lines will appear in the Materials Required But Not Provided LIMITATION The Origin of HIV-1 isolate HTLVIIIB. Nature (1993) 3:363:466-
test line region. Both indicate a positive result. If the specimen does • Specimen collection containers 1. The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid 9.
not contain HIV-1, Subtype O, and/or HIV-2 antibodies, no colored • Lancets (for fingerstick whole blood only) Test Device (Whole Blood/Serum/Plasma) is for in vitro 3. Greenberg AE, Wiktor SZ, DeCock KM, Smith P, Jaffe HW
line will appear in the test line region indicating a negative result. To • Centrifuge diagnostic use only. This test should be used for the detection and Dondero TJ Jr. HIV-2 and natural protection against HIV-1
serve as a procedural control, a colored line will always appear in • Timer of antibodies to HIV in human whole blood, serum or plasma. infection. Science (1996) 272:1959-1960.
the control line region indicating that proper volume of specimen has • Heparinized capillary tubes and dispensing bulb (for fingerstick Neither the quantitative value nor the rate of increase in HIV 4. Travers K, Mboup S, Marlink R, Gueye-Nidaye A, Siby T, Thior
been added and membrane wicking has occurred. whole blood only) antibody concentration can be determined by this qualitative I, Traore I, Dieng-Sarr A, Sankale JL and Mullins C. Natural
test. protection against HIV-1 infection provided by HIV-2. Science
PRECAUTIONS DIRECTIONS FOR USE 2. The HIV 1/2/O Tri-line Human Immunodeficiency Virus Rapid (1995) 268:1612-1615.
• For professional in vitro diagnostic use only. Do not use after Allow the test device, specimen, buffer and/or controls to reach room Test Device (Whole Blood/Serum/Plasma) will only indicate the 5. Arya SK, Beaver B, Jagodzinski L, Ensoli B, Kanki PJ,Albert
expiration date. temperature (15-30°C) prior to testing. presence of antibodies to HIV in the specimen and should not J, Fenyo EM, Biberfeld G, Zagury JF and Laure F. New
• Wear protective clothing such as laboratory coats, disposable 1. Remove the test device from the foil pouch and use it as be used as the sole criteria for the diagnosis of HIV-1, HIV-2, human and simian HIV-related retroviruses possess functional
gloves and eye protection when specimens are being tested. soon as possible. Best results will be obtained if the assay is and/or Subtype O infection. transactivator (tat) gene. Nature (1987) 328:548-550.
• Do not eat, drink or smoke in the area where the specimens or performed within one hour. 3. For confirmation, further analysis of the specimens should be 6. Caetano JA. Immunologic aspects of HIV infection. Acta Med
kits are handled. 2. Place the test device on a clean and level surface. performed, such as ELISA and/or Western Blot analysis. Port (1991) 4 Suppl 1:52S-58S.
• Handle all specimens as if they contain infectious agents. For Serum or Plasma specimens: Hold the dropper vertically 4. As with all diagnostic tests, all results must be interpreted
Observe established precautions against microbiological and transfer 1 drop of serum or plasma (approximately 25 together with other clinical information available to the
hazards throughout testing and follow the standard procedures μL) to the specimen well (S) of the test device, then add 1 physician. ORDERING INFORMATION
for proper disposal of specimens. drop of buffer (approximately 40 μL) and start the timer. See 5. This test is intended for screening purposes only. Results
• The used test should be discarded according to local CATALOG NO. QUANTITY
illustration below. For Venipuncture Whole Blood specimens: should not be used to determine the serotype of HIV infections.
regulations. Hold the dropper vertically and transfer 2 drops of whole blood 6. Due to possible cross reactivity, the appearance of lines in both 1168 001 25 test
• Humidity and temperature can adversely affect results. (approximately 50 μL) to the specimen well (S) of the test T1 and T2 does not necessarily indicate co-infection from HIV- 1168 002 50 test
device, then add 2 drops of buffer (approximately 80 μL) and 1, HIV-2 and Subtype O nor can it identify the serotype.
288 289
Malaria Antigen Test (Whole MATERIALS: Note
Optimal assay performance requires strict adherence to the assay
References
1. Leonard K. Basco, Frederique Marquet, Michael M. Makler, and
Blood) Materials Provided: procedure described in this instruction sheet and any deviations
from the procedure may lead to aberrant results.
Jacques Le Bras : Plasmodium falciparum and Plasmodium
vivax: Lactate Dehydrogenase Activity and its Application for in
Test Device
vitro Drug Susceptibility Assay. Experimental Parasitology 80,
For Plasmodium falciparum & vivax Assay Buffer 260-271 (1995)
Sample Dropper DIRECTIONS FOR USE
2. David L. Vander, Jagt, Lucy A. Hunsaker and John E.
1. Add 5μl of whole blood into sample well (“S” small well)
Heidrich: Partial Purification and Characterisation of Lactate
REF: 1150 001 25 Card Test Materials Not Provided: (Do not use excess blood). Dehydrogenase from Plasmodium falciparum. Molecular and
REF: 1150 002 50 Card Test 5μl Pipette 2. Add two drops or 60-80 μLs of assay buffer into developer well Biochemical Parasitology, 4 (1981) 255-264.
(“A”). 3. David J. Bzik, Barbara A, Fox and Kenneth Gonyer :
SPECIMEN COLLECTION & PREPARATION 3. Read the test result at 20 mins. Expression of Plasmodium falciparum Lactate Dehydrogenase
in Escherichia coli Molecular and Biochemical Parasitology, 59
(1993) 155-166
INTENDED USE (Collection by venipuncture) INTERPRETATION OF RESULTS
For the rapid qualitative determination of Malaria P.falciprum & P. 1. Collect whole blood into a collection tube containing EDTA, 1. P. falciparum Positive reaction 4. Cameron R. Dunn, Mark J. Banfield, John J. Barker,
vivax specific lactate dehydrogenase (pLDH) in human blood for the citrate or heparin) by venipuncture. The presence of two color bands (C and 1) indicates a positive Christopher W. Highm, Kathleen M. Moreton, Dilek Turgut-
Balik, R. Leo Brady and J. John Holbrook. The Structure of
diagnosis of Malaria infection. 2. If specimens are not immediately tested, they should be result for P.falciparum. Lactate Dehydrogenase from Plasmodium falciparum reveals
refrigerated at 2~8°C. For storage periods greater than three a new target for anti-malarial design. Nature Structural Biology
Background days, freezing is recommended. They should be brought to 2. P.vivax Positive reaction 3(11) 1996, 912-915
Malaria is a serious parasitic disease characterized by fever, chills room temperature prior to use. Using the specimen after long- The presence of two color bands (C and 2) indicates a positive 5. Howard, RJ, et al. The secretion of a Malaria Histidine-rich
and anemia and is caused by a parasite that is transmitted from term storage more than three days can cause non-specific result for P. vivax. The pLDH present in the sample reacts with Protein (Pf HRP-2) from Plasmodium falciparum-infected
one human to another by the bite of infected Anopheles mosquitoes. reaction. the pan anti-pLDH conjugate and moves through the test strip Erythrocytes. J. Cell Biol., (1986) 103, 1269-1277
There are four kinds of malaria that can infect humans: Plasmodium 3. When stored at 2~8°C, the whole blood sample should be used where the pLDH is captured by Ovale specific anti-pLDH. 6. Rock, EP, et al. Comparative Analysis of Plasmodium
falciparum, P. vivax, P. Ovale, and P. Malaria. In humans, the within three days. falciparum Histidine-Rich Protein - 2, HRP-I, HRP-II, and HRP-
III in Malaria Parasitology Diverse Origin. Parasitol., (1987)
parasites (called sporozoites) migrate to the liver where they mature 3. Negative reaction
95:209-27.
and release another form, the merozoites. The disease now occurs (Collection using a lancet) The presence of only one band within the result window
in more than 90 countries worldwide, and it is estimated that there 1. Clean the area to be lanced with an alcohol swab. indicates a negative result. 7. Panna, ME, et al. Identification of Plasmodium falciparum
Histidine-Rich Protein - 2 in the of Humans with Malaria. J. Clin.
are over 500 million clinical cases and 2.7 Million malaria-caused 2. Squeeze the end of the fingertip and pierce with a sterile lancet Microbiological 29:1629-1634.
deaths per year. At present, malaria is diagnosed by looking for the provided. 4. Invalid
8. Rodriguez-del Valle, M. et al, Detecting Antigens and Antibodies
parasites in a drop of blood. Blood will be put into a Microscope slide 3. Wipe away the first drop of blood with The test is invalid if the C line does not appear. If this occurs,
in the Urine with Plasmodium falciparum Malaria. Slit Microbiol.,
and stained so that the parasites will be visible under a Microscope. sterile gauze or cotton. the test should be repeated using a new cassette. (1991) 29:1236-1242
4. Take a sample pipette provided, and
Assay Principle while gently squeezing the tube,
Spectrum Malaria Antigen Test contains a membrane strip, which immerse the open end in the blood drop
is pre-coated with two monoclonal antibodies as two separate lines and then gently release the pressure to
across a test strip. One monoclonal antibody (test line 1) is specific draw blood into the sample pipette up to
to the P. vivax (pLDH) and another monoclonal antibody (test line 2) the black line.
is specific to (pLDH) of P. Ovale. Conjugate pad is dispensed with
monoclonal antibodies conjugated to the colloidal gold, which are 1) Gently squeeze 2) Immerse open 3) Gently release to
specific to P. vivax & P. Ovale (pLDH). the tube end in blood draw blood
LIMITATIONS
The Spectrum Malaria Antigen Test is designed for the differential 1. The test procedure, precautions and interpretation of results for
diagnosis between Plasmodium falciparum and Plasmodium Vivax. this test must be followed when testing.
2. Anti-coagulants. heparin, EDTA, and citrate do no affect the
Precautions and Warnings test result
For professional in vitro diagnostic use only. Do not use after 3. This test kit detects Plasmodium lactate dehydrogenase in
expiration date. l Do not eat, drink or smoke in the area where the patient whole blood and is useful as a screening procedure for
specimens or kits are handled . l Handle all specimens as if they malaria diagnosis.
contain infectious agents. Observe established precautions against Directions for use: 4. Do not mix reagent of different lots.
microbiological hazards throughout testing and follow the standard Add whole blood (5 ml) Add 2 drops or 60-80 ml Read Result 5. The test is limited to the detection of antigen of Malaria
procedures for proper disposal of specimens. Into sample well “S” of assay buffer to well “A” in 20 mins. Plasmodium vivax & ovale. Although the test is very accurate
in detecting pLDH, a low incidence of false results can occur.
Wear protective clothing such as laboratory coats, disposable gloves Other clinically available tests. are required if questionable
and eye protection when specimens are being tested. results are obtained. As with all diagnostic tests, a definitive
Humidity and temperature can adversely affect results. clinical diagnosis should not be based on the results of a single
test, but should only be made by the physician after all clinical
STORAGE & STABILITY and laboratory findings have been evaluated
The kit can be stored at room temperature or refrigerated (4-30°C).
The test device is stable through the expiration date printed on the
sealed pouch. The test device must remain in the sealed pouch until
use. DO NOT FREEZE. Do not use beyond the expiration date.
290 291
S.typhi-S.paratyphi “A” Reagents To confirm serum results: The use of a stool sample is Sensitivity:
The test cassette contains anti-salmonella conjugated to gold recommended if serum is used first and a negative result is Spectrum-Diagnostics S.typhi-S.paratyphi assay was run using
Direct Antigen Detection particles and anti-salmonella coated on the membrane. obtained and typhoid is still suspected A second test run on a serum and stool samples versus culture positive samples and found
stool sample should be performed. to give positive results in all cases.
Material Provided: Test Cassette, Droppers, 25 ml Phosphate
REF: 1154 001 25 Card Test Buffer Saline and Instruction for Use 2. Results are then read in as little as10 minutes for strong
REF: 1154 002 50 Card Test positives or up to 20 minutes for weaker positives and to make
Storage and Stability sure negatives are confirmed.
Spectrum-Diagnostics S.typhi-S.paratyphi test kit is stable at
any room temperature between 8 – 30 °C when in the unopened Reading the Test Results
pouches. Negative POSITIVE
Introduction DO NOT FREEZE the kit or expose to temperature extremes. Only one pink/
Typhoid fever is a life threatening illness caused by the bacterium Stability of the kit is 24 months from the date of manufacture – dating purple band
Salmonella typhi, and was observed by Eberth (1880) in the is indicated on the kit label. appears in the C
mesenteric nodes and spleen of fatal cases of typhoid fever. It is (Control) area of
the test card. S. typhi S. paratyphi S.typhi & Paratyphi
common in developing countries where it affects about 12.5 million Precautions
persons annually. The infection is acquired typically by ingestion. On • The test is for In Vitro Diagnostic Use only.
reaching the gut, the bacilli attach themselves to the epithelial cells of • Appropriate infection control and handling procedures should
the intestinal villi and penetrate to the lamina and submucosa. They be followed – do not smoke, eat or drink in the area where the
are then phagocytosed there by polymorphs and macrophages. The test is to be performed. Use suitable clothing and gloves when
ability to resist intracellular killing and to multiply within these cells handling samples and when performing the test.
is a measure of their virulence. They enter the mesenteric lymph • Do not pipette any samples or reagents by mouth.
nodes, where they multiply and, via the thoracic duct, enter the blood • All materials should be considered as potentially infectious. To
stream. A transient bacteremia follows, during which the bacilli are disinfect, either autoclave materials at 121.5°C for 1 hour or
seeded in the liver, gall bladder, spleen, bone marrow, lymph nodes treat with Sodium hypochlorite (1 percent solution). Indeterminate
and kidneys, where further multiplication takes place. Towards the • Do not use test beyond the expiration date indicated. If only one band appears in the 1 or 2 – Test area, or no band
end of the incubation period, there occurs a massive bacteremia appears at all in the C well – Control area. It is then recommended
from these sites, heralding the onset of the clinical symptoms. Sample Collection that a fresh device be used and the test repeated carefully following
The diagnosis of typhoid consists of isolation of the bacilli and the Spectrum-Diagnostics S.typhi-S.paratyphi test can be run on stool the directions in this insert.
demonstration of antibodies. The isolation of the bacilli is very time or serum samples.
consuming and antibody detection is not very specific. Other tests The test works best on fresh samples. If testing cannot be done Quality Control
include the Widal reaction. has developed a test that takes only 10- immediately, they should be stored at 2-8’C after collection for up to A known positive and negative control should be run to ensure
20 minutes and requires only a small quantity of stool or one drop 3 days. If testing cannot be done within 3 days, serum can be stored proper performance. All controls should be handled in the same
of serum* to perform. It is the easiest and most specific method for frozen at -20 °C or colder. manner as patient samples.
detecting S.typhi-S.paratyphi infection. Shipment of samples should comply with local regulations for
transport of etiologic agents. Limitations of the Test
Principle The instructions for use and reading of the test instructions must be
Spectrum-Diagnostics S.typhi-S.paratyphi rapid test is a qualitative Procedure followed carefully for the test to perform properly.
one step immunochromatographic assay. The test employs a 1. Remove as many test cards from the pouches as needed. Lay The Spectrum-Diagnostics S.typhi-S.paratyphi test is designed
conation of monoclonal antibody/colloidal gold dye conjugate and on a clean flat surface. to detect S.typhi-S.paratyphi antigen in stool or serum samples.
a polyclonal antibody immobilized on the solid phase. This will Testing of any other body fluids has not been validated and may not
selectively identify S.typhi-S.paratyphi antigen associated with For stool samples: Add about 0.5 gm stool to approximately yield appropriate results.
typhoid infection with a high degree of sensitivity and specificity. 1 ml of Phosphate Buffered Saline PBS. Mix well and allow to For samples that test positive (reactive) by the Spectrum-Diagnostics
sit for 5 minutes or so to allow the large particles to settle. S.typhi-S.paratyphi test, more specific confirmatory testing should
As the specimen flows through the absorbent pad in the sample Then add 200 ml from the upper layer of the extract to the well be done. A clinical evaluation of the patient’s situation and history
well and through the antibody/colloidal gold complex any S.typhi-S. (A). should also be made before a final diagnosis is established. The use
paratyphi antigen present in the sample binds to the conjugate of a rapid test alone is not sufficient to diagnose S.typhi-S.paratyphi
forming an antigen/antibody complex. The sample and dye complex For serum samples: Using the pipette, add 100 ml of serum/ infection even if antigen is present. Also, a negative result does not
continue to migrate along the membrane to the immobilized plasma to the sample well (A) preclude the possibility of infection with S.typhi-S.paratyphi.
monoclonal antibody. In the presence of S.typhi-S.paratyphi, the
monoclonal antibody captures the complex. This forms a visible The amount of S.typhi-S.paratyphi antigens present in serum Performance Characteristics
pink/purple band in the (B) or test area of the card. If no antigen is typically less than that in stool. This may decrease the
is present, there is no line formation in the (B) area. The remaining sensitivity of the test when using serum depending how soon Specificity:
complex continues to migrate to another immobilized antibody on after the onset of the infection the test is performed. Early The antibodies used in the Spectrum-Diagnostics S.typhi-S.
the membrane in the (C) or Control area of the card, and is captured infection typically exhibits greater levels of the antigen in the paratyphi assay were developed specifically against Salmonella
which then forms a band indicating proper performance of the test. serum than in later infection. typhi and Salmonella paratyphi LPS antigen.
292 293
Syphilis Ab Combo Rapid Test SYMBOLS IN PRODUCT LABELLING
no anticoagulants in Vacutainer®) by veinpuncture.
2. Allow the blood to clot.
2. POSITIVE RESULT: If both C and T bands are developed,
the test indicates for the presence of anti-Tp antibody in the
(Serum / Plasma / Whole Blood) EC REP Authorised Representative Temperature Limitation 3. Separate the serum by centrifugation. specimen. The result is positive.
4. Carefully withdraw the serum into a new pre-labeled tube.
REF: 1204 001 30 test Test specimens as soon as possible after collecting. Store
specimens at 2°C-8°C if not tested immediately. Store specimens
REF Catalogue Number for use at 2°C-8°C up to 5 days. The specimens should be frozen at -20°C
Consult instructions for use Manufactured by for longer storage.
Samples with positive results should be confirmed with alternative
INTENDED USE Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen testing method(s) and clinical findings before a positive
The Spectrum Syphilis Ab Combo Rapid Test is a lateral flow an internal control (C band) which should exhibit a burgundy specimens to room temperature slowly and mix gently. Specimens determination is made.
chromatographic immunoassay for the qualitative detection of colored band of the immunocomplex of goat anti-rabbit IgG/rabbit containing visible particulate matter should be clarified by
antibodies including IgG, IgM, and IgA to Treponema pallidum (Tp) IgG-gold conjugate regardless of color development on the T band. centrifugation before testing. Do not use samples demonstrating 3. INVALID: If no C band is developed, the assay is invalid
in human serum, plasma or whole blood. It is intended to be used Otherwise, the test result is invalid and the specimen must be gross lipemia, gross hemolysis or turbidity in order to avoid regardless of color development on the T band as indicated
as a screening test and as an aid in the diagnosis of infection with retested with another device. interference on result interpretation. below. Repeat the assay with a new device.
Tp. Any reactive specimen with the Spectrum Syphilis Ab Combo Blood
Rapid Test must be confirmed with alternative testing method(s) REAGENTS AND MATERIALS PROVIDED Drops of whole blood can be obtained by either finger tip puncture
and clinical findings. 1. Individually sealed foil pouches containing: or veinpuncture. Do not use any hemolized blood for testing.
a. One cassette device.
SUMMARY AND EXPLANATION OF THE TEST b. One plastic dropper. Whole blood specimens should be stored in refrigeration (2°C-8°C)
Tp, a spirochete bacterium, is the causative agent of the venereal c. One desiccant. if not tested immediately. The specimens must be tested within 24
disease syphilis. Although syphilis rates are declining in the United 2. Sample diluent (1 vial, 5 mL) hours of collection. PERFORMANCE CHARACTERISTICS
States after an epidemic outbreak between 1986 and 1990, the 3. One package insert (instruction for the use). Clinical Performance
incidence of syphilis in Europe has increased since 1992, especially ASSAY PROCEDURE A total of 1055 samples from susceptible subjects were tested by
in the countries of the Russian Federation, where peaks of 263 MATERIALS MAY BE REQUIRED AND NOT PROVIDED Step 1: Bring the specimen and test components to room the Spectrum Syphilis Ab Combo Rapid Test and by a TPHA test.
cases per 100,000 have been reported. In 1995, WHO reported 12 1. Positive Control temperature if refrigerated or frozen. Mix the specimen well prior to Comparison for all subjects is shown in the following table.
million new cases of syphilis. Currently, the positive rate of syphilis 2. Negative Control assay once thawed.
serological tests in HIV-infected individuals has been rising recently. Step 2: When ready to test, open the pouch at the notch and Spectrum Syphilis Ab Combo Rapid Test
MATERIALS REQUIRED BUT NOT PROVIDED remove device. Place the test device on a clean, flat surface. TPHA Positive Negative Total
Serological detection of anti-Tp antibody has been long recognized Step 3: Be sure to label the device with specimen’s ID number.
in the diagnosis of syphilis since the natural course of the infection 1. Clock or Timer Positive 318 0 318
2. Lancing device for whole blood test Step 4: Fill the plastic dropper with the specimen.
was characterized by periods without clinical manifestations. Holding the dropper vertically, dispense 1 drop (about 30-45 µL) of Negative 2 735 737
Both IgM and IgG antibodies were detected in sera from patients serum/plasma or 1 drop of whole blood (about 40-50 µL) into the
with primary and secondary syphilis. The IgM antibody may WARNINGS AND PRECAUTIONS Total 320 735 1055
For in Vitro Diagnostic Use sample well making sure that there are no air bubbles.
be detectable towards the second week of infection, while IgG
1. This package insert must be read completely before performing Immediately add1 drop (about 35-50 µL) of Sample Diluent to the
antibody appears later, at about 4 weeks. These antibodies could Relative Sensitivity: 100%, Relative Specificity: 99.7%, Overall
last for several years or even decades in the serum of a patient with the test. Failure to follow the insert gives inaccurate test results. sample well.
Agreement: 99.8%
untreated latent syphilis. 2. Do not open the sealed pouch, unless ready to conduct the Precision
assay. Within run and between run precisions have been determined
Antigens such as Rapid Plasma Cardiolipin antigen (RPR) and Tp 3. Do not use expired devices. by testing 15 replicates with three of the samples: a negative, a
bacterial extracts have been used in the syphilis serological tests 4. Bring all reagents to room temperature (15°C-30°C) before
for decades. However, RPR antigen is a non-treponema antigen, weak positive, and a strong positive sample. The negative, weaker
use. positive, and strong positive samples were correctly identified in all
derived from bovine heart. Antibody to RPR antigen does not 5. Do not use the components in any other type of test kit as a
develop until 1-4 weeks after the appearance of the chancre, thus of the tests each time.
substitute for the components in this kit.
this antigen lacks of sensitivity to primary syphilis. The Tp extracts 6. Do not use hemolized blood specimen for testing.
are prepared from inoculated rabbit testis and contain a certain LIMITATIONS OF TEST
7. Wear protective clothing and disposable gloves while 1. The Assay Procedure and the Assay Result Interpretation
amount of contaminated materials such as flagella, which can lead
handling the kit reagents and clinical specimens. Wash hands must be followed closely when testing the presence of anti-
to cross reactions with borreliae and leptospires in the serological
test. In addition, the composition of extracts may vary from lot to thoroughly after performing the test. Tp antibody in serum, plasma or whole blood from individual
lot. Recently, several highly immunogenic Tp specific antigens have 8. Users of this test should follow the US CDC Universal subjects. Failure to follow the procedure may give inaccurate
Precautions for prevention of transmission of HIV, HBV and Step 5: Set up timer.
been identified and used as an alternative to the traditional antigens Step 6: Results can be read in 15 minutes. Positive results can results.
with the advantages of high specificity and reproducibility. other blood-borne pathogens. 2. The Spectrum Syphilis Ab Combo Rapid Test is limited to
9. Do not smoke, drink, or eat in areas where specimens or kit be visible in as short as 1minute.
the qualitative detection of anti-Tp antibody in human serum
The Spectrum Syphilis Ab Combo Rapid Test permits the reagents are being handled. or plasma. The intensity of the test band does not linear
measurement of antibodies (IgM, IgG and IgA) to recombinant 10. Dispose of all specimens and materials used to perform the Don’t read result after 15 minutes. To avoid confusion, discard
the test device after interpreting the result. correlation with the antibody titer in the specimen.
antigens of Tp in blood rapidly and reliably without instrumentation. test as biohazardous waste. 3. A negative result for an individual subject indicates absence
11. Handle the Negative and Positive Control in the same manner of detectable anti-Tp antibody. However, a negative test result
as patient specimens. QUALITY CONTROL
1. Internal Control: This test contains a built-in control feature, does not preclude the possibility of exposure to or infection
TEST PRINCIPLE 12. The testing results should be read within 15 minutes after a with Tp.
The Spectrum Syphilis Ab Combo Rapid Test is a lateral flow specimen is applied to the sample well or sample pad of the the C band. The C line develops after adding specimen and
sample diluent. Otherwise, review the whole procedure and 4. A negative result can occur if the quantity of the anti-Tp
chromatographic immunoassay. The test cassette consists of: device. Reading result after 15 minutes may give erroneous antibody present in the specimen is below the detection limits
1) a burgundy colored conjugate pad containing recombinant Tp results. repeat test with a new device.
2. External Control: Good Laboratory Practice recommends of the assay, or the antibodies that are detected are not present
antigens conjugated with colloidal gold (Tp conjugates) and rabbit 13. Do not perform the test in a room with strong air flow, ie. an during the stage of disease in which a sample is collected.
IgG-gold conjugates, 2) a nitrocellulose membrane strip containing electric fan or strong air-conditioning. using external controls, positive and negative, to assure
the proper performance of the assay, particularly under the 5. Some specimens containing unusually high titer of heterophile
a test band (T band) and a control band (C band). The T band is antibodies or rheumatoid factor may affect expected results.
pre-coated with non-conjugated recombinant Tp antigens, and the REAGENT PREPARATION AND STORAGE following circumstances:
a. New operator uses the kit, prior to performing testing of 6. The results obtained with this test should only be interpreted
C band is pre-coated with goat anti-rabbit IgG antibody. INSTRUCTIONS in conjunction with other diagnostic procedures and clinical
All reagents are ready to use as supplied. Store unused test device specimens.
b. A new lot of test kit is used. findings.
unopened at 2°C -30°C. If stored at 2°C -8°C, ensure that the test
device is brought to room temperature before opening. The test c. A new shipment of kits is used.
d. The temperature used during storage of the kit falls REFERENCES
device is stable through the expiration date printed on the sealed 1. Tichonova, L., K. Borisenko, H.Ward, A.meheus, et al.
pouch. Do not freeze the kit or expose the kit over 30°C. outside of 2-30°C.
e. The temperature of the test area falls outside of 15-30°C. Epidemics of syphilis in the Russian Federation: Trends,
f. To verify a higher than expected frequency of positive or origins, and priorities for control. Lancet 1997; 350:210-213.
SPECIMEN COLLECTION AND HANDLING 2. Bailey MJ, Thomas CM, Cockayne A, Strugnell RA, Penn CW.
Consider any materials of human origin as infectious and handle negative results.
g. To investigate the cause of repeated invalid results. Cloning and expression of Treponema pallidum antigens in
When an adequate volume of test specimen is dispensed into the them using standard biosafety procedures. Escherichia coli. J Gen Microbiol 1989; 135 ( Pt 9):2365-78.
sample well of the cassette, the specimen migrates by capillary Plasma 3. Rufli T. Syphilis and HIV infection. Dermatologica 1989;
action across the cassette. Anti-Tp antibody, if present in the 1. Collect blood specimen into a lavender, blue or green INTERPRETATION OF ASSAY RESULT
1. NEGATIVE RESULT: If only the C band is developed, the test 179:113-117.
specimen will bind to the Tp conjugates. The immunocomplex is top collection tube (containing EDTA, citrate or heparin,
then captured on the membrane by the pre-coated Tp antigen, respectively in Vacutainer® ) by veinpuncture. indicates that no detectable anti-Tp antibody is present in the
forming a burgundy colored T band, indicating a Tp antibody 2. Separate the plasma by centrifugation. specimen. The result is negative.
positive test result. 3. Carefully withdraw the serum into a new pre-labeled tube.
Serum
Absence of the T band suggests a negative result. The test contains 1. Collect blood specimen into a red top collection tube (containing
294 295
ToRCH SYMBOLS IN PRODUCT LABELLING
NEGATIVE: One colored line appears in the control line region
(C) of every section. Non-appearance of a visible line in the test
CMV Rapid Test Device vs. EIA
Toxo/Rubella/CMV/HSV 1/2 IgM Antibodies Combo line region (T) of any section is indicative of a negative test result for Method EIA Total
Rapid Test Device (Serum/Plasma) EC REP Authorised Representative Temperature Limitation Results
that specific section, viz. Toxo, Rub, CMV, and HSV 1/2. Results Positive Negative
IVD For in-vitro diagnostic use Use by/Expiration Date INVALID: Control line fails to appear in any section. Insufficient CMV Rapid
Positive 18 5 23
LOT Batch Code/Lot number CAUTION. specimen volume or incorrect procedural techniques are the most Test
REF: 1204 001 25 test likely reasons for control line failure. Review the procedure and Negative 1 461 462
REF Catalogue Number Consult instructions for use
repeat the test with a new test device. If the problem persists, Total Results 19 466 485
A rapid test for the qualitative detection of IgM antibodies to Consult instructions for use Manufactured by
discontinue using the test kit immediately and contact your local
Toxoplasma gondii (Toxo), Rubella virus (Rubella), Cytomegalovirus distributor. Relative Sensitivity: 94.7% (74.0%-99.9%)*
(CMV), and Herpes simplex virus 1/2 (HSV 1/2) in serum or plasma. Relative Specificity: 98.9% (97.5%-99.7%)*
For professional in vitro diagnostic use only. SPECIMEN COLLECTION AND PREPARATION QUALITY CONTROL Relative Accuracy: 98.8% (97.3%-99.5%)*
• The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/ A procedural control is included in the test individually for all the * 95% Confidence Interval
INTENDED USE Plasma) can be performed using either serum or plasma four sections. Four colored lines appearing in control line regions
The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/ specimen. (C) of all four sections is the internal procedural control. It confirms HSV 1/2 Rapid Test Device vs. EIA
Plasma) is a rapid chromatographic immunoassay for the qualitative • Separate the serum or plasma from blood as soon as possible sufficient specimen volume and correct procedural technique.
detection of IgM antibodies to Toxoplasma gondii (Toxo), Rubella to avoid hemolysis. Only clear, non-hemolyzed specimens can Control standards are not supplied with this kit; however, it is Method EIA Total
virus (Rubella), Cytomegalovirus (CMV), and Herpes simplex virus be used. recommended that positive and negative controls be tested as good Results Positive Negative Results
1/2 (HSV 1/2) in serum or plasma to aid in the diagnosis of ToRCH. • Testing should be performed immediately after the specimens laboratory practice to confirm the test procedure and to verify proper HSV 1/2
Positive 12 1 13
have been collected. Do not leave the specimens at room test performance. Rapid Test
SUMMARY temperature for prolonged periods. Specimens may be stored Negative 2 300 302
ToRCH is an acronym for a group of infectious diseases that, at 2-8°C for up to 3 days. For long term storage, specimens LIMITATION Total Results 14 301 315
while infecting the pregnant women, may cause birth defects in should be kept below -20°C. 1. The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/
their newborns.1 ToRCH stands for 4 different infections that can • Bring specimens to room temperature prior to testing. Frozen Plasma) is for in vitro diagnostic use only.This test should be Relative Sensitivity: 85.7% (57.2%-98.2%)*
adversely affect the pregnant women and the fetus, newborn specimens must be completely thawed and mixed well prior used for the detection of IgM antibodies to Toxo, Rubella, Relative Specificity: 99.7% (98.2%-100.0%)*
children including birth defects and often leading to abortion. The to testing. Specimens should not be frozen and thawed CMV and HSV 1/2 in serum or plasma specimens. Neither the Relative Accuracy: 99.0% (97.2%-99.8%)*
four infections are Toxomplasma gondii (A spirochete), Rubella repeatedly. quantitative value nor the rate of increase in the concentration * 95% Confidence Interval
(Virus), CMV - Cytomegalovirus (Virus), HSV 1/2 - Herpes Simplex • If specimens are to be shipped, they should be packed of IgM antibodies to Toxo, Rubella, CMV and HSV 1/2 can be
Virus 1 and/or 2 (Virus). The infections usually cause few, if any, in compliance with local regulations for the transportation determined by this qualitative test. Precision
symptoms in the pregnant woman, but pose greater risks of of etiologic agents. 2. The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/ Intra-Assay
serious birth defects for neonates. Infections caused by ToRCH - Plasma) will only indicate the presence of IgM antibodies to Within-run precision has been determined by using 10 replicates of
Toxoplasma, Rubella Virus, Cytomegalo Virus (CMV) and Herpes MATERIALS Toxo, Rubella, CMV and HSV 1/2 in the specimen and should three specimens containing negative, low positive and high positive
Simplex Virus (HSV) - is the major cause of BOH (Bad Obstetric Materials Provided not be used as the sole criteria for the diagnosis of any of the of Toxo, Rubella, CMV or HSV 1/2. The negative and positive values
History).2 • Test devices ToRCH infections for which the positive result is obtained. were correctly identified >99% of the time
Risks are severe, if the mother gets the infection in the first • Disposable droppers 3. As with all diagnostic tests, all results must be considered with
trimester as the baby’s organs start to form in this stage. General • Buffer other clinical information available to the physician. Inter-Assay
symptoms include premature birth, growth retardation, neurological • Package insert 4. If the test result is negative and clinical symptoms persist, Between-run precision has been determined by using the same
abnormalities, damage of the eye, liver, heart and ear as well as bone additional follow-up testing using other clinical methods three specimens of negative, low positive and high positive of
lesions. Microcephaly, hydrocephaly, seizures and psychomotor Materials Required But Not Provided is suggested. A negative result for any one out of the four Toxo, Rubella, CMV and HSV 1/2 in 3 independent assays. Three
retardation accompany these malformations. • Specimen collection container infections of ToRCH at any time does not preclude the different lots of the ToRCH IgM Antibodies Combo Rapid Test Device
The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/ • Centrifuge (for plasma only) possibility of that particular infection. (Serum/Plasma) have been tested using negative, low positive and
Plasma) is a rapid chromatographic immunoassay for the qualitative • Timer high positive specimens. The specimens were correctly identified
detection of IgM antibodies to Toxo, Rubella, CMV, and HSV 1/2 in EXPECTED VALUES >99% of the time.
serum or plasma specimens. DIRECTIONS FOR USE The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/
Allow the test device, specimen, and/or controls to equilibrate to Plasma) has been compared with leading commercial EIA Cross-Reactivity
PRINCIPLE room temperature (15-30°C) prior to testing. Toxo, Rubella, CMV, and HSV 1/2 tests, demonstrating an overall Sera containing known amounts of antibodies to Toxo, Rubella,
The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/ 1. Bring the pouch to room temperature before opening it. accuracy of 98.4% for Toxo, 99.1% for Rubella, 98.8% for CMV, and CMV and HSV 1/2 have been tested with HIV, HCV, SYP, HBV, and
Plasma) is a qualitative, lateral flow immunoassay for the detection Remove the test device from the sealed pouch and useit as 99.0% for HSV 1/2. RF. No cross-reactivity was observed, indicating that the ToRCH
of IgM antibodies to Toxo, Rubella, CMV, and HSV 1/2 in serum or soon as possible. Best results will be obtained if the assay is IgM Antibodies Combo Rapid Test Device (Serum/Plasma) has a
plasma specimens. In this test, antigens of Toxo, Rub, CMV and performed within one hour. PERFORMANCE CHARACTERISTICS high degree of specificity for IgM antibodies to Toxo, Rubella, CMV,
HSV 1/2 are coated in the test line regions of each section in the 2. Place the test device on a clean and level surface. Hold the Sensitivity and Specificity and HSV 1/2.
test. During testing, the serum or plasma specimen reacts with Goat dropper vertically and transfer 1 full drop of serum or plasma The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/
anti-human IgM coated particles in the test strip. The mixture then (approximately 10 µL) and 2 drops of buffer (approximately 80 Plasma) was compared with leading commercial Interfering Substances
migrates upward on the membrane by capillary action and reacts µL) to each specimen well of the test device respectively, and EIA Toxo, Rubella, CMV, and HSV 1/2 tests, the results show that The ToRCH IgM Antibodies Combo Rapid Test Device (Serum/
with the Toxo, Rub, CMV and HSV then start the timer. Avoid trapping air bubbles in the specimen the ToRCH IgM Antibodies Combo Rapid Test Plasma) has been tested and no interference was observed
1/2 specific antigens on the membrane in the test line regions of well. See illustration below. Device (Serum/Plasma) has a high sensitivity and specificity for in specimens containing 110 mg/mL human albumin, 1 mg/mL
the respective sections. The presence of a colored line in the test 3. Wait for the colored line(s) to appear. The results should each of its sections. bilirubin, 10 mg/mL hemoglobin, 0.2mg/mL cholesterol and 15 mg/
line region of a particular section indicates a positive result for the be read at 15 minutes. Do not interpret the results after 20 mL triglycerides.
corresponding infection, viz. Toxo, Rub, CMV, HSV 1/2, while its minutes. Toxo Rapid Test Device vs. EIA The following compounds have also been tested using the ToRCH
absence indicates a negative result for that infection. To serve IgM Antibodies Combo Rapid Test Device (Serum/Plasma) and no
as a procedural control, a colored line will always appear in the Method EIA Total interference was observed
respective control line regions of all the four strips indicating that Results
proper volume of specimen has been added and membrane wicking Results Positive Negative Acetaminophen Acetylsalicylic Ascorbic Acid Bilirubin
Toxo
has occurred. Rapid Test Positive 54 12 66 Acid
Caffeine Ethanol EDTA Glucose
REAGENTS Negative 3 846 849
The test contains Goat anti-human IgM coated particles and Toxo Gentisic acid Phenothiazine Phenylpropanolamine Salicylic
antigens, Rub antigens, CMV antigens, and HSV 1/2 antigens Total Results 57 858 915 Acid
coated on the membrane.
Relative Sensitivity: 94.7% (85.4%-98.9%)*
PRECAUTIONS Relative Specificity: 98.6% (97.6%-99.3%)*
• For professional in vitro diagnostic use only. Do not use after Relative Accuracy: 98.4% (97.3%-99.1%)* BIBLIOGRAPHY
INTERPRETATION OF RESULTS * 95% Confidence Interval 1. S M Kadri, Torch Test: Test & Inference, INDIAN JOURNAL
expiration date. (Please refer to the illustration above)
• Do not eat, drink or smoke in the area where the specimens OF THE PRACTISING DOCTOR, January 2005, Vol. I, No.
POSITIVE: Rubella Rapid Test Device vs. EIA 4 : P16-18
or kits are handled. Toxo Positive: *Two colored lines appear in the ‘Toxo’ Section. One
• Do not use test if the package is damaged. 2. Rajendra B Surpam, Usha P Kamlakar, RK Khadse, MS
line should be in the control line region Method EIA Total Qazi, & Suresh V Jalgaonkar, Serological study for TORCH
• Handle all specimens as if they contain infectious agents. (C) and another line should be in the test line region (T).
Observe established precautions against microbiological Results Positive Negative Results infections in women with bad obstetric history, The Journal of
Rubella Positive: *Two colored lines appear in the ‘Rub’ Section. Rubella Obstetrics and Gynecology of India, January/February 2006,
hazards throughout testing and follow the standard procedures One line should be in the control line region (C) and another line Positive 19 2 21
for proper disposal of specimens. Rapid Test Vol. 56, No. 1 : P 41-43
should be in the test line region (T). Negative 1 298 299
• Wear protective clothing such as laboratory coats, disposable
gloves and eye protection when specimens are being tested. CMV Positive: *Two colored lines appear in the ‘CMV’ Section. Total Results 20 300 320
• Humidity and temperature can adversely affect results. One line should be in the control line region
• The used test should be discarded according to local Relative Sensitivity: 95.0% (75.1%-99.9%)* CATALOG NO. QUANTITY
(C) and another line should be in the test line region (T). Relative Specificity: 99.3% (97.6%-99.9%)*
regulations. HSV 1/2 Positive: *Two colored lines appear in the ‘HSV 1/2’ Relative Accuracy: 99.1% (97.3%-99.8%)* 1204 001 25 test
Section. One line should be in the control line region (C) and * 95% Confidence Interval
STORAGE AND STABILITY another line should be in the test line region (T).
Store as packaged in the sealed pouch either at room temperature *NOTE: The intensity of the color in test line regions (T) will vary
or refrigerated (2-30°C). The test device is stable through the depending on the concentrations of IgM antibodies present in the
expiration date printed on the sealed pouch. The test device must specimen. Therefore, any shade of color in test line regions (T)
remain in the sealed pouch until use. DO NOT FREEZE. Do not use should be considered positive.
beyond the expiration date.
296 297
Toxo IgG/IgM Rapid Test- SYMBOLS IN PRODUCT LABELLING
assay, once thawed.
Step 2: When ready to test, open the pouch at the notch and
3. INVALID: If no C line is developed, the assay is invalid
regardless of any burgundy color in the test bands as indicated
remove device. Place the test device on a clean, flat surface. below. Repeat the assay with a new device.
Step 3: Be sure to label the device with the specimen’s ID
(Serum / Plasma) number.Step 4: Fill the plastic dropper with the specimen.
LOT Batch Code/Lot number CAUTION. Consult instructions Holding the dropper vertically, dispense 1 drop (about 30-45 µL) of
REF Catalogue Number for use specimen into the sample well.
REF: 1196 001 30 test Then add 1 drop (about 30-45 µL) of Sample Diluent immediately.
INTENDED USE
The Spectrum Toxo IgG/IgM Rapid Test is a lateral flow MATERIALS REQUIRED BUT NOT PROVIDED
chromatographic immunoassay for the simultaneous detection and 1. Clock or Timer PERFORMANCE CHARACTERISTICS
differentiation of IgG and IgM anti-Toxoplasma gondii (T. gondii) 1. Clinical Performance For IgM Test
in human serum or plasma. This kit is intended to be used as a WARNINGS AND PRECAUTIONS A total of 302 samples from susceptible subjects were tested by
screening test and as an aid in the diagnosis of infection with T. For in Vitro Diagnostic Use the Spectrum Toxo IgG/IgM Rapid Test and by a commercial IgM
gondii. Any reactive specimen with the Spectrum Toxo IgG/IgM 1. This package insert must be read completely before performing EIA kit. Comparison of the results for all subjects is shown in the
Rapid Test must be confirmed with alternative testing method(s) the test. Failure to follow the insert gives inaccurate test results. Step 5: Set up timer. following table.
and clinical findings. 2. Do not open the sealed pouch unless ready to conduct the Step 6: Results can be read in 15 minutes. Positive results can
assay. be visible in as short as 1 minute. Spectrum Toxo IgG/IgM Rapid Test
SUMMARY AND EXPLANATION OF THE TEST 3. Do not use expired devices. IgM EIA Positive Negative Total
T. gondii is an obligate intracellular protozoan parasite with a 4. Bring all reagents to room temperature (15°C-30°C) before Don’t read result after 15 minutes. To avoid confusion, discard
worldwide distribution. Serological data indicates that approximately use. the test device after interpreting the result. Positive 2 0 2
30% of the population of most industrialized nations is chronically 5. Do not use the components in any other type of test kit as a
substitute for the components in this kit. QUALITY CONTROL Negative 2 298 300
infected with the organism.
6. Do not use hemolized blood for testing. 1. Internal Control: This test contains a built-in control feature, Total 4 298 302
A variety of serological tests for antibodies to T. gondii have been 7. Wear protective clothing and disposable gloves while the C line. The C line develops after adding the specimen and
used as an aid in diagnosis of acute infection and to assess previous handling the kit reagents and clinical specimens. Wash hands the sample diluent. If the C line does not develop, review the Relative Sensitivity: 100% , Relative Specificity: 99.3%, Overall
exposure to the organism. These tests are: the Sabin-Feldman thoroughly after performing the test. whole procedure and repeat the test with a new device. Agreement: 99.3%
dye test, direct agglutination, indirect hemagglutination, latex 8. Users of this test should follow the US CDC Universal 2. External Control: Good Laboratory Practice recommends
agglutination, indirect immunofluorescence and ELISA. Recently, Precautions for prevention of transmission of HIV, HBV and using external controls, positive and negative, to assure 2. Clinical Performance For IgG Test
lateral flow chromatographic immunoassay such as the Spectrum other blood-borne pathogens. the proper performance of the assay, particularly under the A total of 324 samples from susceptible subjects were tested by
Toxo IgG/IgM Rapid Test has been introduced to the clinic for the 9. Do not smoke, drink or eat in areas where specimens or kit following circumstances: the Spectrum Toxo IgG/IgM Rapid Test and by a commercial IgG
instant detection of T. gondii infection. reagents are being handled. a. New operator uses the kit, prior to performing the testing EIA kit. Comparison of the results for all subjects is shown in the
10. Dispose of all specimens and materials used to perform the of specimens. following table.
TEST PRINCIPLE test as bio-hazardous waste. b. A new lot of test kits is used.
The Spectrum Toxo IgG/IgM Rapid Test is a lateral flow 11. Handle the negative and positive control in the same manner c. A new shipment of kits is used. Spectrum Toxo IgG/IgM Rapid Test
chromatographic immunoassay. The test cassette consists of: 1) a as patient specimens. d. The temperature used during storage of the kits fall
12. The test results should be read within 15 minutes after IgG EIA Positive Negative Total
burgundy colored conjugate pad containing recombinant T. gondii outside of 2-30°C.
antigens conjugated with colloidal gold (T. gondii conjugates) and a specimen is applied to the sample well or sample pad of e. The temperature of the test area falls outside of 15-30°C. Positive 22 2 24
rabbit IgG-gold conjugates, 2) a nitrocellulose membrane strip the device. Reading the results after 15 minutes may give f. To verify a higher than expected frequency of positive or
containing two test bands (M and G bands) and a control band (C erroneous results. negative results. Negative 3 297 300
band). The M band is pre-coated with monoclonal anti-human IgM 13. Do not perform the test in a room with strong air flow, i.e. an g. To investigate the cause of repeated invalid results. Total 25 299 324
for detection of IgM anti-T. gondii antibody, G band is pre-coated electric fan or strong air-conditioning.
with reagents for detection of IgG anti-T.gondii antibody, and the C INTERPRETATION OF ASSAY RESULT Relative Sensitivity: 91.6% , Relative Specificity: 99.0%, Overall
band is pre-coated with goat anti-rabbit IgG. REAGENT PREPARATION AND STORAGE 1. NEGATIVE RESULT: If only the C line is present, the Agreement: 98.5%
INSTRUCTIONS absence of any burgundy color in both the test lines (M and
All reagents are ready to use as supplied. Store unused test devices G) indicates that no anti-T. gondii antibodies are detected in
unopened at 2°C-30°C. Do not freeze the kit. Do not expose the
LIMITATIONS OF TEST
the specimen. The result is negative or non-reactive 1. The Assay Procedure and the Interpretation of Assay Result
kit over 30°C. The positive and negative controls should be kept at
sections must be followed closely when testing for the
2°C-8°C or the temperature indicated. If stored at 2°C-8°C, ensure
presence of antibodies to T. gondii in serum or plasma from
that the test device is brought to 15°C-30°C before opening. The
individual subjects. Failure to follow the procedure may give
test device is stable through the expiration date printed on the
inaccurate results.
When an adequate volume of test specimen is dispensed into the sealed pouch.
2. The Spectrum Toxo IgG/IgM Rapid Test is limited to the
sample well of the test cassette, the specimen migrates by capillary 2. POSITIVE RESULT: qualitative detection of antibodies to T. gondii in human serum
action across the cassette. IgM anti-T. gondii if present in the SPECIMEN COLLECTION AND HANDLING a. In addition to the presence of the C line, if only the M or plasma. The intensity of the test line does not have a linear
specimen will bind to the T. gondii conjugates. The immunocomplex Consider any materials of human origin as infectious and handle line is developed, the test indicates the presence of IgM correlation with the antibody titer in the specimen.
is then captured on the membrane by the pre-coated anti-human them using standard bio-safety procedures. anti-T. gondii in the specimen. The result is positive or 3. A negative result for an individual subject indicates absence
IgM antibody forming a burgundy colored M line, indicating a T. Plasma reactive. of detectable anti-T. gondii antibodies. However, a negative
gondii IgM positive or reactive test result. 1. Collect blood specimen into a lavender, blue or green
test result does not preclude the possibility of exposure to or
top collection tube (containing EDTA, citrate or heparin,
infection with T. gondii.
IgG anti-T. gondii if present in the specimen will bind to the T. gondii respectively in Vacutainer®) by veinpuncture.
4. A negative result can occur if the quantity of the anti-T. gondii
conjugates. The immunocomplex is then captured by the pre- 2. Separate the plasma by centrifugation.
antibodies present in the specimen is below the detection limits
coated reagents on the membrane forming a burgundy colored G 3. Carefully withdraw the plasma into new pre-labeled tube.
of the assay or the antibodies that are detected are not present
line, indicating a T. gondii IgG positive or reactive test result. Serum b. In addition to the presence of the C line, if only the G during the stage of disease in which a sample is collected.
1. Collect blood specimen into a red top collection tube (containing line is developed, the test indicates the presence of IgG 5. Some specimens containing unusually high titers of heterophile
Absence of any T lines (M and G) suggests a negative or non- no anticoagulants in Vacutainer®) by veinpuncture. anti-T. gondii in the specimen. The result is positive or antibodies or rheumatoid factor may affect expected results.
reactive result. The test contains an internal control (C band) which 2. Allow the blood to clot. reactive. 6. If the symptoms persist when the result from Spectrum
should exhibit a burgundy colored line of the immunocomplex of 3. Separate the serum by centrifugation.
Toxo IgG/IgM Rapid Test is negative or non-reactive, it is
goat anti-rabbit IgG/rabbit IgG-gold conjugate regardless of color 4. Carefully withdraw the serum into a new pre-labeled tube.
recommended to re-sample the patient a few days later or test
development on any of the T lines. Otherwise, the test result is with an alternative test device.
invalid and the specimen must be retested with another device. Test specimens as soon as possible after collecting. Store
specimens at 2°C-8°C if not tested immediately.
REAGENTS AND MATERIALS PROVIDED Store specimens at 2°C-8°C for up to 5 days. The specimens should c. In addition to the presence of the C line, if both the M and
be frozen at -20°C for longer storage. findings.
1. Individually sealed foil pouches containing: the G lines are developed, the test indicates the presence
a. One cassette device Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen of both IgG and IgM anti-T. gondii in the specimen. The
b. One desiccant specimens to room temperature slowly and mix gently. Specimens result is also positive or reactive. REFERENCES
2. Plastic droppers containing visible particulate matter should be clarified by 1. Pyndiah N, Krech U, Price P and Wilhelm J: Simplified
3. Sample diluent (1 bottle, 5 mL) centrifugation before testing. Do not use samples demonstrating chromatographic separation of immunoglobulin M from G and
4. One package insert (instruction for use) gross lipemia, gross hemolysis or turbidity in order to avoid its application to Toxoplasma indirect immunofluorescence. J.
interference on result interpretation. Clin. Micro.1979, 9:170-174
MATERIALS MAY BE REQUIRED AND NOT PROVIDED 2. Fraser KB, Shirodaira PV, and Stanford CF: Fluorescent
1. Positive Control ASSAY PROCEDURE Samples with positive results should be confirmed with staining and human IgM Br. Med. J. 1971, 3:707
2. Negative Control Step 1: Bring the specimen and test components to room alternative testing method(s) and clinical findings before a 3. Montoya JG, Rosso F. Diagnosis and management of
temperature if refrigerated or frozen. Mix the specimen well, prior to positive determination is made. toxoplasmosis. Clin Perinatol. 2005, 32(3):705-26.
298 299
Troponin-I SYMBOLS IN PRODUCT LABELLING
PERFORMANCE CHARACTERISTICS
Troponin I Test Device EC REP Authorised Representative Temperature Limitation
Sensitivity:
(Serum/Plasma/Whole Blood) IVD For in-vitro diagnostic use Use by/Expiration Date The Spectrum Troponin I Test Device can detect cTnI in serum or
LOT Batch Code/Lot number CAUTION. Consult instructions plasma with concentration of 1.0 ng /mL or greater.
REF: 1172 001 25 test Catalogue Number for use
REF
REF: 1172 002 50 test Interference Substances:
A rapid test for the qualitative detection of Cardiac Troponin I in Levels of the following substances do not appear to interfere with
Human serum, plasma or whole blood. the Cardiac Troponin I Assay.
For professional in vitro test use only.
Human Albumin 16 g/dL
INTENDED USE STORAGE AND STABILITY QUALITY CONTROL Bilirubin (unconjugated) 60 mg/dL
For the rapid qualitative determination of Cardiac troponin I (cTnI) Store as packaged in the sealed pouch at 4-30 °C. The test device Free Hemoglobin 4 g/dL
Using individual Spectrum Troponin I Test Device cassettes as
in human whole blood, serum and plasma as an aid in the diagnosis is stable through the expiration date printed on the sealed pouch. Triglycerides 1,300 mg/dL
described in the Assay Procedure above, run 1 Positive Control
of myocardial infarction The test device must remain in the sealed pouch until use. Do not
and 1 Negative Control (provided upon request) under the following
freeze. Method Comparison
circumstances to monitor test performance:
SUMMARY 1. A new operator uses the kit, prior to performing testing of Serum samples (n=121) collected from individuals after being
Troponin I (TnI) is part of the troponin complex which, together with SPECIMEN COLLECTION AND PREPARATION specimens. admitted to a hospital emergency department with chest pain.
tropomyosin, forms the main component that regulates the Ca+2- Whole blood, plasma or serum may be used as samples for 2. A new test kit is used. The samples were tested with the Troponin I test and with FDA
sensitive ATP-ase activity of actomyosin in striated muscle (skeletal this procedure. Collect blood in a tube containing heparin as 3. A new shipment of kits is used. approved cardiac troponin I test kit. The correlation between the
and cardiac). The troponin complex consists of three subunits, the anticoagulant. Guidelines recommended by the National 4. The temperature used during storage of the kit falls outside of tests is shown below:
troponin T(TnT), troponin I(TnI), and troponin C (TnC). Each subunit Committee for Clinical Laboratory Standards (NCCLS) should 2°C-30°C. FDA approved Troponin I test
has a distinct function with TnC as the site of Ca+2 binding, TnT be followed when collecting, transporting and processing patient 5. The temperature of the test area falls outside of 15°C-30°C. Positive Negative Total
the tropomyosin binding, and TnI as the inhibitory subunit. Different samples. Since cTnI is relatively unstable, it is recommended that
isoforms of TnI exist in the skeletal and cardiac muscles (sTnI and fresh samples be used as soon as possible to collect critical patient Spectrum Positive 31 1 32
cTnI, respectively) with distinct immunologic epitopes that allow the information. Heat inactivation of samples may lead to hemolysis or
INTERPRETATION OF RESULTS Negative 1 88 89
production of cardiac-specific TnI antibodies. The cardiac marker, protein denaturation and therefore should be avoided.
Invalid: Total 32 89 121
troponin I has been established as useful tools in the diagnosis of Whole blood samples should be tested within 2 hours of collection. If
A distinctive colored band in the control window should always
acute myocardial infarction (AMI). Troponin I is found in blood at specimens are to be shipped, they should be packed in compliance
appear. If no pink band is present in the Control window within 15 Comparative Sensitivity: 96.9 % (31/32)
elevated concentrations approximately 4- 6 hours after the onset with federal regulations covering the transportation of etiologic
minutes, the test is invalid, and the sample should be run again with Comparative Specificity: 98.9% (88/89)
of chest pain and peak at 12-24 hours. Troponin I levels remain agents.
a new test device. Overall Agreement: 98.35 % (119/121)
elevated for up to 14 days. The use of this marker is an aid in the MATERIALS
diagnosis of AMI after myocardial infarction. Materials Provided Negative result:
BIBLIOGRAPHY
• Test device(s) A color band will appear only in the control window,
PRINCIPLE 1. Hamm, C.W., et al. Emergency room triage of patients with
• Instructions for use which indicates a negative result.
The Troponin I Rapid Test Device employs a solid-phase acute chest pain by means of rapid testing for cardiac troponin
chromatographic immunoassay technology to qualitatively detect Materials Required But Not Provided T or troponin I. New Eng.J.Med.337:1648 (1997).
Positive result: 2. Mehegan, J.P. And Tobacman, L.S. Cooperative interaction
the elevation of troponin I in human blood samples. When a • Vacutainer tube containing heparin as an anticoagulant
Two bands will appear in the control and test window, which between troponin molecules bound to the cardiac thin filament.
sample of blood is dispensed into the sample well, red blood cells • Timer
indicates a positive result. J.Biol.Chem. 266:966 (1991).
are removed by the built in separation system. Troponin I in the • Positive and Negative Controls
specimen makes a complex with the specific dye conjugate and • Calibrated Pipette 3. Antman, E.M., et al. Cardiac-specific troponin I levels to predict
Notes for Result Interpretation: the risk of mortality inpatients with acute coronary syndromes.
biotinylated anti-troponin I antibody. This complex migrates through • The color intensity of the Test and Control bands may increase
the test area containing immobilized streptavidin. The antibody dye- DIRECTIONS FOR USE New Eng.J.Med. 335:1342 (1996).
beyond 15 minutes. As the membrane in the reading window 4. Ebashi,S., Ca2+ and the contractile proteins. J.Mol.Cell.
troponin I-biotinylated antibody complex bind to the immobilized 1. Open the foil pouch, remove the Cardiac test device, and
dries up, the color intensity of the bands and background Cardiol. 16:129 (1984).
streptavidin in the Test area. Unbound dye complexes migrate out lay the device on a level surface. Label the device with the
change and thus may interfere with reading the test results. 5. Bodor, S.G. et al., Development of monoclonal antibody for an
of the Test area and are later captured in the Control area. Visible patient’s name or control number.
• For best results, the test result should be read at 15 minutes. assay of cardiac troponin I and preliminary results in suspected
pinkish-purple bands will appear in the Test and Control areas if the 2. Using a micropipette, add 80 ml of whole blood or 60 ml of
The result, particularly a result which is negative before 15 cases of myocardial infarction. Clin.Chem. 38:2203 (1992).
concentrations of troponin I is above established cutoff values. If serum or plasma specimen into the Sample well.
minutes, should not be read beyond 15 minutes. 6. Ellis, A.K. Serum protein measurements and the diagnosis of
the troponin I concentration in the specimen is 0.6 ng/ml or greater, 3. Read the test result in 15 minutes.
• The test bands will appear before the control band in most acute myocardial infarction. Circulation 83:1107 (1991).
a band is present in the troponin I area. If a band is present only in strong positive cases. The test bands may be darker than the
the Control area, the test result is read as negative, indicating that NOTES:
control band.
the Troponin I concentrations are all below the cutoff values. If no • If the test has been stored in the refrigerator, allow it to
• The test bands may appear after the control band in weak
band is present in the Control area, the test is invalid and another return to room temperature before testing. Do not open
positive cases, and the test band may be weaker than the
test must be run, regardless of the presence or absence of band(s) the foil pouch until you are ready to perform the test. ORDERING INFORMATION
control band.
in the Test Area. • When the specimen is dispensed, do not position the
pipette tip too high from the device’s Sample area, in CATALOG NO. QUANTITY
order to prevent the samples from splashing. LIMITATION
PRECAUTIONS 1172 001 25 test
The results of the Cardiac TnI Assay are to be used in conjunction
• For in vitro diagnostic use only. 1172 002 50 test
with other clinical information such as clinical signs and symptoms
• Do not use beyond the expiration date. and other test results to diagnose myocardial infarction. A negative
• Use separate syringe or clean pipette tips for different result obtained from a patient’s sample 16 hours after the onset of
specimens. Do not pipette by mouth. chest pain may help in ruling out AMI. A positive assay result from a
• Do not smoke, eat or drink in areas in which specimens or kits patient suspected of AMI may be used as an indicative of myocardial
are handled. damage and requires further confirmation. Serial sampling of
• Wear disposable gloves while handling specimens and running patients suspected of AMI is also recommended due to the delay
the tests, and thoroughly wash hands afterwards. between the onset of symptoms and the release of protein markers
• All patient samples should be handled as if they are capable into the bloodstream.Samples containing an unusually high titer of
of transmitting diseases. Observe established good laboratory certain antibodies, such as human anti mouse or human anti goat
procedures for proper disposal of specimens, used pipette tips antibodies, may affect the performance of the test.
or syringes, and used test devices.
• The test device should remain in its sealed pouch until ready
EXPECTED VALUES
for use.
The Cardiac Troponin I Assay is designed to yield a positive result
• Humidity and temperature can adversely affect results.
for cTnI concentrations at or more than 0.6 ng/mL
300 301
Troponin-I SYMBOLS IN PRODUCT LABELLING
LIMITATION
The results of the Cardiac TnI Assay are to be used in conjunction
Troponin I Test Device EC REP Authorised Representative Temperature Limitation with other clinical information such as clinical signs and symptoms
(Serum/Plasma/Whole Blood) IVD For in-vitro diagnostic use Use by/Expiration Date and other test results to diagnose myocardial infarction. A negative
LOT Batch Code/Lot number CAUTION. Consult instructions result obtained from a patient’s sample 16 hours after the onset of
chest pain may help in ruling out AMI. A positive assay result from a
REF
REF: 1173 001 30 test patient suspected of AMI may be used as an indicative of myocardial
damage and requires further confirmation. Serial sampling of
patients suspected of AMI is also recommended due to the delay
A rapid test for the qualitative detection of Cardiac Troponin I in between the onset of symptoms and the release of protein markers
Human serum, plasma or whole blood. into the bloodstream.Samples containing an unusually high titer of
For professional in vitro test use only. certain antibodies, such as human anti mouse or human anti goat
antibodies, may affect the performance of the test.
INTENDED USE STORAGE AND STABILITY
Store as packaged in the sealed pouch at 4-30 °C. The test device EXPECTED VALUES
For the rapid qualitative determination of Cardiac troponin I (cTnI)
is stable through the expiration date printed on the sealed pouch. The Cardiac Troponin I Assay is designed to yield a positive result
in human whole blood, serum and plasma as an aid in the diagnosis
The test device must remain in the sealed pouch until use. Do not for cTnI concentrations at or more than 0.6 ng/mL
of myocardial infarction
freeze.
SUMMARY PERFORMANCE CHARACTERISTICS
Troponin I (TnI) is part of the troponin complex which, together with SPECIMEN COLLECTION AND PREPARATION
Whole blood, plasma or serum may be used as samples for Sensitivity:
tropomyosin, forms the main component that regulates the Ca+2-
sensitive ATP-ase activity of actomyosin in striated muscle (skeletal this procedure. Collect blood in a tube containing heparin as
The Spectrum Troponin I Test Device can detect cTnI in serum or
and cardiac). The troponin complex consists of three subunits, the anticoagulant. Guidelines recommended by the National
plasma with concentration of 1.0 ng /mL or greater.
troponin T(TnT), troponin I(TnI), and troponin C (TnC). Each subunit Committee for Clinical Laboratory Standards (NCCLS) should
has a distinct function with TnC as the site of Ca+2 binding, TnT be followed when collecting, transporting and processing patient
Interference Substances:
the tropomyosin binding, and TnI as the inhibitory subunit. Different samples. Since cTnI is relatively unstable, it is recommended that
isoforms of TnI exist in the skeletal and cardiac muscles (sTnI and fresh samples be used as soon as possible to collect critical patient Levels of the following substances do not appear to interfere with
cTnI, respectively) with distinct immunologic epitopes that allow the information. Heat inactivation of samples may lead to hemolysis or the Cardiac Troponin I Assay.
production of cardiac-specific TnI antibodies. The cardiac marker, protein denaturation and therefore should be avoided.
troponin I has been established as useful tools in the diagnosis of Whole blood samples should be tested within 2 hours of collection. If QUALITY CONTROL Human Albumin 16 g/dL
acute myocardial infarction (AMI). Troponin I is found in blood at specimens are to be shipped, they should be packed in compliance Using individual Spectrum Troponin I Test Device cassettes as Bilirubin (unconjugated) 60 mg/dL
elevated concentrations approximately 4- 6 hours after the onset with federal regulations covering the transportation of etiologic described in the Assay Procedure above, run 1 Positive Control Free Hemoglobin 4 g/dL
of chest pain and peak at 12-24 hours. Troponin I levels remain agents. and 1 Negative Control (provided upon request) under the following Triglycerides 1,300 mg/dL
elevated for up to 14 days. The use of this marker is an aid in the circumstances to monitor test performance:
diagnosis of AMI after myocardial infarction. 1. A new operator uses the kit, prior to performing testing of Method Comparison
MATERIALS
specimens. Serum samples (n=121) collected from individuals after being
Materials Provided
2. A new test kit is used. admitted to a hospital emergency department with chest pain.
PRINCIPLE • Test device(s)
3. A new shipment of kits is used. The samples were tested with the Troponin I test and with FDA
The Troponin I Rapid Test Device employs a solid-phase • Sample diluent (1 x 5 mL)
4. The temperature used during storage of the kit falls outside of approved cardiac troponin I test kit. The correlation between the
chromatographic immunoassay technology to qualitatively detect • One plastic dropper.
2°C-30°C. tests is shown below:
the elevation of troponin I in human blood samples. When a • Instructions for use
5. The temperature of the test area falls outside of 15°C-30°C. FDA approved Troponin I test
sample of blood is dispensed into the sample well, red blood cells
Materials Required But Not Provided
are removed by the built in separation system. Troponin I in the Positive Negative Total
• Vacutainer tube containing heparin as an anticoagulant
specimen makes a complex with the specific dye conjugate and
• Timer INTERPRETATION OF RESULTS Positive 31 1 32
biotinylated anti-troponin I antibody. This complex migrates through Spectrum
• Positive and Negative Controls Invalid:
the test area containing immobilized streptavidin. The antibody dye- Negative 1 88 89
• Calibrated Pipette A distinctive colored band in the control window should always
troponin I-biotinylated antibody complex bind to the immobilized
streptavidin in the Test area. Unbound dye complexes migrate out appear. If no pink band is present in the Control window within 15 Total 32 89 121
of the Test area and are later captured in the Control area. Visible ASSAY PROCEDURE minutes, the test is invalid, and the sample should be run again with
pinkish-purple bands will appear in the Test and Control areas if the 1. Bring the specimen and test components to room temperature a new test device. Comparative Sensitivity: 96.9 % (31/32)
concentrations of troponin I is above established cutoff values. If if refrigerated or frozen. Mix the specimen well prior to assay Comparative Specificity: 98.9% (88/89)
the troponin I concentration in the specimen is 0.6 ng/ml or greater, once thawed. Negative result: Overall Agreement: 98.35 % (119/121)
a band is present in the troponin I area. If a band is present only in 2. When ready to test, open the pouch at the notch and remove A color band will appear only in the control window,
the Control area, the test result is read as negative, indicating that device. Place the test device on a clean, flat surface. which indicates a negative result. BIBLIOGRAPHY
the Troponin I concentrations are all below the cutoff values. If no 3. Be sure to label the device with specimen’s ID number. 1. Hamm, C.W., et al. Emergency room triage of patients with
band is present in the Control area, the test is invalid and another 4. For whole blood test Positive result: acute chest pain by means of rapid testing for cardiac troponin
test must be run, regardless of the presence or absence of band(s) Apply 1 drop of whole blood (about 40-50 µL) into the sample Two bands will appear in the control and test window, which T or troponin I. New Eng.J.Med.337:1648 (1997).
in the Test Area. well. indicates a positive result. 2. Mehegan, J.P. And Tobacman, L.S. Cooperative interaction
Then add 1 drop (about 35-50 µL) of Sample Diluent between troponin molecules bound to the cardiac thin filament.
immediately. For serum or plasma test Notes for Result Interpretation: J.Biol.Chem. 266:966 (1991).
PRECAUTIONS
Fill the pipette dropper with the specimen. • The color intensity of the Test and Control bands may increase 3. Antman, E.M., et al. Cardiac-specific troponin I levels to predict
• For in vitro diagnostic use only.
Holding the dropper vertically, dispense 1 drop (about 30-45 beyond 15 minutes. As the membrane in the reading window the risk of mortality inpatients with acute coronary syndromes.
• Do not use beyond the expiration date.
µL) of specimen into the sample well making sure that there dries up, the color intensity of the bands and background New Eng.J.Med. 335:1342 (1996).
• Use separate syringe or clean pipette tips for different
are no air bubbles. change and thus may interfere with reading the test results. 4. Ebashi,S., Ca2+ and the contractile proteins. J.Mol.Cell.
specimens. Do not pipette by mouth.
Then add 1 drop (about 35-50 µL) of Sample Diluent • For best results, the test result should be read at 15 minutes. Cardiol. 16:129 (1984).
• Do not smoke, eat or drink in areas in which specimens or kits
immediately The result, particularly a result which is negative before 15 5. Bodor, S.G. et al., Development of monoclonal antibody for an
are handled.
5. Set up timer. minutes, should not be read beyond 15 minutes. assay of cardiac troponin I and preliminary results in suspected
• Wear disposable gloves while handling specimens and running
6. Results can be read in 15 minutes. Positive results can be • The test bands will appear before the control band in most cases of myocardial infarction. Clin.Chem. 38:2203 (1992).
the tests, and thoroughly wash hands afterwards.
visible in as short as 1 minute. strong positive cases. The test bands may be darker than the 6. Ellis, A.K. Serum protein measurements and the diagnosis of
• All patient samples should be handled as if they are capable
of transmitting diseases. Observe established good laboratory control band. acute myocardial infarction. Circulation 83:1107 (1991).
NOTES: • The test bands may appear after the control band in weak
procedures for proper disposal of specimens, used pipette tips
• If the test has been stored in the refrigerator, allow it to positive cases, and the test band may be weaker than the
or syringes, and used test devices. ORDERING INFORMATION
return to room temperature before testing. Do not open control band.
• The test device should remain in its sealed pouch until ready
the foil pouch until you are ready to perform the test. CATALOG NO. QUANTITY
for use.
• When the specimen is dispensed, do not position the
• Humidity and temperature can adversely affect results. 1173 001 30 test
pipette tip too high from the device’s Sample area, in
order to prevent the samples from splashing.
302 303
Typhoid IgG/IgM Rapid Test- SYMBOLS IN PRODUCT LABELLING
PERFORMANCE CHARACTERISTICS
1. Clinical Performance For IgM Test
A total of 334 samples from susceptible subjects were tested by the
Spectrum Typhoid IgG/IgM Rapid Test and by a commercial S. typhi
IgM EIA. Comparison for all subjects is shown in the following table.
(Serum / Plasma)
Spectrum Typhoid IgG/IgM Rapid Test
REF: 1200 001 30 test
Consult instructions for use Manufactured by IgM EIA Positive Negative Total
Positive 31 3 34
INTENDED USE MATERIALS REQUIRED BUT NOT PROVIDED Step 5: Set up timer.
The Spectrum Typhoid IgG/IgM Rapid Test is a lateral flow 1. Clock or Timer Step 6: Results can be read in 15 minutes. Negative 2 298 300
immunoassay for the qualitative detection and differentiation of IgG Total 33 302 334
and IgM anti-Salmonella typhi (S. typhi) and paratyphi in human WARNINGS AND PRECAUTIONS Don’t read result after 15 minutes. To avoid confusion, discard
serum or plasma. It is intended to be used as a screening test and For in Vitro Diagnostic Use the test device after interpreting the result.
Relative Sensitivity: 91%, Relative Specificity: 99.3%, Overall
as an aid in the diagnosis of infection with S. typhi and paratyphi. 1. This package insert must be read completely before performing Agreement: 98.5%
Any reactive specimen with the Spectrum Typhoid IgG/IgM Rapid the test. Failure to follow the insert gives inaccurate test results. QUALITY CONTROL
Test must be confirmed with alternative testing method(s). 2. Do not open the sealed pouch, unless ready to conduct the 1. Internal Control: This test contains a built-in control feature,
the C band. The C line develops after adding specimen and 2. Clinical Performance For IgG Test
assay. A total of 314 samples from susceptible subjects were tested by
SUMMARY AND EXPLANATION OF THE TEST 3. Do not use expired devices. sample diluent. Otherwise, review the whole procedure and
repeat test with a new device. the Spectrum Typhoid IgG/IgM Rapid Test and by a commercial
Typhoid fever and paratyphi fever are bacterial infections caused 4. Bring all reagents to room temperature (15°C-30°C) before S. typhi IgG EIA kit. Comparison for all subjects is shown in the
by Salmonella typhi and paratyphi A, B, and C respectively, which use. 2. External Control: Good Laboratory Practice recommends
using external controls, positive and negative, to assure following table.
are transmitted through the ingestion of tainted food and water. 5. Do not use the components in any other type of test kit as a
Worldwide an estimated 17 million cases and 600,000 associated substitute for the components in this kit. the proper performance of the assay, particularly under the
following circumstances: Spectrum Typhoid IgG/IgM Rapid Test
deaths occur annually1. Patients who are infected with HIV are at 6. Do not use hemolized blood specimen for testing.
significantly increased risk of clinical infection. 1-5% of patients 7. Wear protective clothing and disposable gloves while a. New operator uses the kit, prior to performing testing of IgG EIA Positive Negative Total
become chronic carriers harboring S. typhi in the gallbladder. handling the kit reagents and clinical specimens. Wash hands specimens.
thoroughly after performing the test. b. A new lot of test kit is used. Positive 13 1 14
The clinical diagnosis of infections depends on isolation of S. typhi 8. Users of this test should follow the US CDC Universal c. A new shipment of kits is used.
d. The temperature used during storage of the kit falls Negative 2 298 300
and paratyphi from blood, bone marrow or a specific anatomic Precautions for prevention of transmission of HIV, HBV and
lesion. In facilities that can not afford to perform this complicated other blood-borne pathogens. outside of 2-30°C. Total 15 299 314
and time-consuming procedure, Filix-Widal test is used to facilitate 9. Do not smoke, drink, or eat in areas where specimens or kit e. The temperature of the test area falls outside of 15 -30°C.
diagnosis. However, many limitations lead to difficulties in the reagents are being handled. f. To verify a higher than expected frequency of positive or Relative Sensitivity: 92.9% , Relative Specificity: 99.3%, Overall
interpretation of the Widal test3,4. 10. Dispose of all specimens and materials used to perform the negative results. Agreement: 99.0%
test as biohazardous waste. g. To investigate the cause of repeated invalid results.
In contrast, the Spectrum Typhoid IgG/IgM Rapid Test is a simple, 11. Handle the Negative and Positive Control in the same manner 3. Performance comparison with blood culture
fast laboratory test that simultaneously detects and differentiates as patient specimens. INTERPRETATION OF ASSAY RESULT Nine (9) S. paratyphi A and eleven (11) S. typhi specimens
IgG and IgM antibodies to S. typhi and paratyphi antigen5 thus 12. The testing results should be read within 15 minutes after a 1. NEGATIVE OR NON-REACTIVE RESULT: If only the C band confirmed with the blood culture were tested with the Spectrum
aiding in the determination of current or previous exposure to S. specimen is applied to the sample well or sample pad of the is present, the absence of any burgundy color in the both test Typhoid IgG/IgM Rapid Test. The Spectrum Typhoid IgG/IgM Rapid
typhi and paratyphi. IgM positive or IgM /IgG both positive suggest device. Reading the test after 15 minutes may give erroneous bands (M and G) indicates that no anti-S. typhi or paratyphi Test correctly indentified 9 S. paratyphi A and 10 S. typhi specimens.
current infection, while IgG positive suggests late stage of infection, results. antibody is detected in the specimen. The result is negative The agreement was 95%.
previous infection, or latent infection. 13. Do not perform the test in a room with strong air flow, ie. an or non-reactive.
electric fan or strong air-conditioning. LIMITATIONS OF TEST
TEST PRINCIPLE 1. The Assay Procedure and the Test Result Interpretation must
The Spectrum Typhoid IgG/IgM Rapid Test is a lateral flow REAGENT PREPARATION AND STORAGE be followed closely when testing the presence of antibodies
chromatographic immunoassay. The test cassette consists of: 1) a INSTRUCTIONS to S. typhi or paratyphi in serum or plasma from individual
burgundy colored conjugate pad containing recombinant H antigen All reagents are ready to use as supplied. Store unused test devices subjects. Failure to follow the procedure may give inaccurate
and O antigen conjugated with colloidal gold (HO conjugates) and unopened at 2°C-30°C. If stored at 2°C-8°C, ensure that the test 2. POSITIVE OR REACTIVE RESULT: results.
rabbit IgG-gold conjugates, 2) a nitrocellulose membrane strip device is brought to room temperature before opening. The test a. In addition to the presence of C band, if only M band is 2. The Spectrum Typhoid IgG/IgM Rapid Test is limited to the
containing two test bands (G and M bands) and a control band device is stable through the expiration date printed on the sealed developed, the test indicates for the presence of anti- S. qualitative detection of antibodies to S. typhi or paratyphi
(C band). The M band is pre-coated with monoclonal anti-human pouch. Do not freeze the kit or expose the kit over 30°C. typhi or paratyphi IgM in the specimen. The result is IgM in human serum or plasma. The intensity of the test band
IgM for the detection of IgM anti-S. typhi and paratyphi, G band is positive or reactive. does not have linear correlation with the antibody titer in the
pre-coated with reagents for the detection of IgG anti-S. typhi and SPECIMEN COLLECTION AND HANDLING specimen.
paratyphi , and the C band is pre-coated with goat anti rabbit IgG. Consider any materials of human origin as infectious and handle 3. A negative result for an individual subject indicates absence
them using standard biosafety procedures. of detectable anti-S. typhi or paratyphi antibodies. However,
Plasma a negative test result does not preclude the possibility of
1. Collect blood specimen into a lavender, blue or green exposure to S. typhi or paratyphi .
top collection tube (containing EDTA, citrate or heparin, b. In addition to the presence of C band, if only G band is 4. A negative result can occur if the quantity of anti-S. typhi or
respectively in Vacutainer® ) by veinpuncture. developed, the test indicates for the presence of anti- S. paratyphi antibodies present in the specimen is below the
2. Separate the plasma by centrifugation. typhi or paratyphi IgG in the specimen. The result is IgG detection limit of the assay, or the antibodies that are detected
3. Carefully withdraw the plasma into a new pre-labeled tube. positive or reactive. are not present during the stage of disease in which a sample
Serum is collected.
When an adequate volume of test specimen is dispensed into 1. Collect blood specimen into a red top collection tube 5. If the symptom persists, while the result from Spectrum
the sample well of the cassette, the test specimen migrates by (containing no anticoagulants in Vacutainer®) by veinpuncture. Typhoid IgG/IgM Rapid Test is negative or non-reactive result,
capillary action across the test cassette. IgM antibodies if present 2. Allow the blood to clot. it is recommended to re-sample the patient few days late or
in the patient specimen will bind to the HO conjugates. The 3. Separate the serum by centrifugation. test with an alternative test method, such as bacterial culture
immunocomplex is then captured on the membrane by the pre- 4. Carefully withdraw the serum into a new pre-labeled tube. c. In addition to the presence of C band, both M and G method.
coated anti-human IgM antibody, forming a burgundy colored M bands are developed, the test indicates for the presence 6. Some specimens containing unusually high titer of heterophile
band, indicating a S. typhi or paratyphi IgM positive test result. Test specimens as soon as possible after collecting. Store of anti-S. typhi or paratyphi IgG and IgM in the specimen. antibodies or rheumatoid factor may affect expected results.
specimens at 2°C-8°C if not tested immediately. The result is both IgG and IgM positive or reactive. 7. The results obtained with this test should only be interpreted
IgG antibodies if present in the patient specimen will bind to the in conjunction with other diagnostic procedures and clinical
HO conjugates. The immunocomplex is then captured by the pre- Store specimens at 2°C-8°C for up to 5 days. The specimens should
be frozen at -20°C for longer storage. findings.
coated reagents on the membrane, forming a burgundy colored G
band, indicating a S. typhi or paratyphi IgG positive test result. Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen REFERENCES
specimens to room temperature slowly and mix gently. Specimens 1. Gotuzzo E, Frisancho O, Sanchez J, Liendo G, Carillo C,
Absence of any test bands (M and G) suggests a negative result. Samples with positive or reactive results should be confirmed Black RE, Morris JG. Association between the acquired
containing visible particulate matter should be clarified by with alternative testing method(s) and clinical findings before
The test contains an internal control (C band) which should exhibit centrifugation before testing. Do not use samples demonstrating immunodeficiency syndrome and infection with Salmonella
a burgundy colored band of the immunocomplex goat anti-rabbit a positive determination is made. typhi or Salmonella paratyphi in an endemic typhoid area.
gross lipemia, gross hemolysis or turbidity in order to avoid
IgG/rabbit IgG-gold conjugate regardless of the color development interference on result interpretation. Archives of Internal Medicine 1991; 151: 381-2.
on any of the test bands. Otherwise, the test result is invalid and the 3. INVALID: If no C band is developed, the assay is invalid 2. Ivanoff BN, Levine MM, Lambert PH. Vaccination against
specimen must be retested with another device. regardless of any burgundy color in the test bands as indicated typhoid fever: present status. Bulletin of the World Health
ASSAY PROCEDURE below. Repeat the assay with a new device. Organization 1994; 72: 957-71.
REAGENTS AND MATERIALS PROVIDED Step 1: Bring the specimen and test components to room 3. Ismail A, Hai OK, Kader ZA. Demonstration of an antigenic
1. Individually sealed foil pouches containing: temperature if refrigerated or frozen. Mix the specimen well prior to protein specific for Salmonella typhi. Biochem Biophys Res
a. One cassette device. assay once thawed. Commun. 1991;181(1):301-5.
b. One desiccant. Step 2: When ready to test, open the pouch at the notch and 4.
2. Plastic droppers. remove device. Place the test device on a clean, flat surface. 5. Pang T. False positive Widal test in nontyphoid Salmonella
3. Sample Diluent (1 bottle, 5 mL) Step 3: Be sure to label the device with specimen’s ID number. infection. Southeast Asian Journal of Tropical Medicine and
4. One package insert (instruction for use). Step 4: Fill the plastic dropper with the specimen. Public Health 1989; 20: 163-4.
Holding the dropper vertically, dispense 1 drop (about 30-45 mL)
MATERIALS MAY BE REQUIRED AND NOT PROVIDED of specimen into the sample well making sure that there are no air
1. Positive Control bubbles.
2. Negative Control Then add 1 drop (about 35-50 µL) of Sample Diluent immediately.
304 305
URI-TRAK SYMBOLS IN PRODUCT LABELLING pH: The pH value of fresh urine of healthy people varies between
EC REP Authorised Representative Temperature Limitation pH 5 and pH 6. The colour scale gives a clear distinction of pH value
Test strips for rapid determination of Glucose, Ketones, Protein and IVD For in-vitro diagnostic use Use by/Expiration Date between pH 5 and pH 9.
pH-value in Urine
Consult instructions for use Manufactured by o
Store at room temperature (15 to 30 C) out of direct sunlight.
REF: 1100 010 (Uri-trak 1) for Glucose Do not use after expiration date.
REF: 1100 020 (Uri-trak 2) for Glucose & Ketones
REF: 1100 030 (Uri-trak 3) for Glucose, Protein & pH Precautions
The Kit contain a non-poisonous and harmless desiccant. In case
this desiccant is swallowed accidently, then drink plenty of water.
Intended Use Instructions for use Disposal

Spectrum Uri-trak is a screening test strips for detection of glucose, Dip the test strip for approximately 1 second into the fresh urine. Please dispose all used dipsticks in accordance with your local laws
ketones, protein and pH-value in urine. Certain configuration of strips Draw it across the rim of the container to remove excess urine. After and regulations.
may be read instrumentally, using the appropriate Urine Chemistry 30 to 60 seconds compare the test strip with the colour scale. The
Analyzers. best time for ncomparison is after 30 seconds. Colour changes that
take place after more than 2 minutes are of no significance. When
Principle tested the urine should not be older than 2 hours. ORDERING INFORMATION
Glucose: The detection is based on the glucoseoxidase-peroxidase- CATALOG NO QUANTITY
chromogen reaction. Apart from glucose, no other compound in Evaluation – Sources of Error
1100 010
urine is known to give a positive reaction. Glucose: Pathological glucose concentrations are indicated by a 1100 020 100 Test Strip for each
colour change from green to bluish green. Yellow or greenish test 1100 030
Ketones: The test is based on the principle of Legal’s test. Acetoacetic fields should be considered negative or normal. The colour fields
acid and acetone form with sodium nitroprusside in alkaline medium correspond to the following ranges of glucose concentrations:
a violet coloured complex. neg. (yellow), neg. or normal (greenish), 50, 150, 500 and 1000 mg/
dl or neg. (yellow), neg. or normal (greenish), 2.8, 8.3, 27.8 and 55.5
Protein: The test is based on the „protein error“ principle of indicators. mmol/l
The test zone is buffered to a constant pH value and changes colour The influence of ascorbic acid (vitamin C) has been largely
from yellow to greenish blue in the presence of albumin. Other eliminated. An inhibitory effect is produced by gentisic acid. Falsely
proteins are indicated with less sensitivity. positive reactions can be produced by a residue of peroxide
containing cleansing agents.
pH: The test paper contains indicators which clearly change colour
between pH 5 and pH 9 (from orange to green to turquoise). Ketones: The test is more sensitive to acetoacetic acid than to
acetone. Values of 10 mg/dl acetoacetic acid or 50 mg/dl acetone are
Reactive Ingredients indicated. The colour fields correspond to the following acetoacetic
acid values:
Glucose: 0 (negative), 25(+), 100(++) and 300(+++) mg/dl or
2% w/w glucose oxidase; 1% w/w peroxidase; 10% w/w TMB; 70% 0 (negative), 2.5(+), 10(++) and 30(+++) mmol/l
w/w Buffer; 17% w/w nonreactive ingredients. Phenylketones in higher concentrations interfere with the test, and
will produce variable colours.
Ketones: b-Hydroxybutyric acid is not detected. Phthalein compounds
5% w/w sodium nitroprusside; 95% w/w Buffer. interfere by producing a red colouration.
Protein: Protein: The minimum sensitivity of the test strip is 10 mg protein/

0.2% w/w tetrabromophenol blue; 97.4% w/w Buffer; 2.4% w/w dl urine. The colour fields correspond to the following ranges of
nonreactive ingredients. albumin concentrations:
negative, 30, 100 and 500 mg/dl or negative, 0.3, 1.0 and 5.0 g/l
pH: Falsely positive results are possible in alkaline urine samples (pH L
0.2% w/w methyl red; 2.8% w/w bromothymol blue; 97% w/w 9), after infusions with polyvinylpyrrolidone (blood substitute), after
nonreactive ingredients. intake of medicaments containing quinine and also by disinfectant
residues in the urine sampling vessel. The protein colouration may
Specimen Collection be masked by the presence of medical dyes (e.g. methylene blue)
Use a fresh urine samples that is less than 2 hours old and place it or beetroot pigments.
in a clean, dry container. Do not centrifuge.
The presence of usual urine preservatives will not affect the test
results.
306 307
URI-TRAK-10 Specific Gravity (SG): High-buffered alkaline urine may cause
1 2 3
SYMBOLS IN PRODUCT LABELLING diminished result, whereas high-buffered acidic urine may cause
REAGENT STRIPS FOR URINALYSIS EC REP Authorised Representative Temperature Limitation slightly elevated result.
IVD For in-vitro diagnostic use Use by/Expiration Date Expected values
Batch Code/Lot number CAUTION. Consult instructions Blood: Normally, no hemoglobin is detectable in urine (0.010mg/dl;
LOT
REF: 1100 100 (Uri-trak -10) for Ten Parameters Catalogue Number for use
3 RBC/µl). When hemoglobin appears in urine it indicates kidney
REF
disease or a urinary tract disorder. Blood may often be found in the
urine of menstruating females.
Bilirubin: Normally no bilirubin is detectable in urine by even
the most sensitive methods. Even trace amounts of bilirubin are
Summary and Explanation Specific Gravity (SG): Ionic solutes present in the urine cause Quality control sufficiently abnormal to require further investigation.
URI-TRAK-10 Reagent Strips are dip-and-read test strips for In protons to be released from a polyelectrolyte. As the protons For best results, performance of reagent strips should be confirmed
Vitro Diagnostic Use only for testing ten parameters in urine. Test are released the pH decreases and produces a color change of by testing known negative and positive specimen or controls Urobilinogen: The normal urobilinogen range is 0.1 to 1.0 Ehrlich
result may provide information regarding the status of carbohydrate bromothymol blue from blue-green to yellow-green. whenever a new bottle is first opened. Each laboratory should unit /dl. If results exceed the concentration of 2.0 mg/dl, the patient
metabolism, kidney and liver function, acid-base balance, and Ingredients: bromthymol blue 4.8%W/W establish its own goals for adequate standards of performance. and the urine specimen should be evaluated further.
urinary tract infection. It is measured by comparison of test paper poly(methyl vinyl ether-alt- maleic anhydride) Each lab worker should ensure that it complies with government Ketone: Ketone bodies should not be detected in normal urine
attached to a plastic strip with the color chart blocks printed on the 90.2%W/W and local requirements. specimens with this reagent.
vial label. The strips may be read visually. They can also be read
instrumentally, using urine chemistry analyzers. Storage and Handling Limitations of procedure Glucose: A small amounts of glucose(up to 30mg/dl) are normally
Store in a cool, dry place at temperatures between 2OC ~ 30OC. As with all laboratory tests, definitive diagnostic or therapeutic excreted by the kidney.
Principle Do not store the strips in a refrigerator or freezer. Store away decisions should not be based on any single result of method. Protein: Normal urine specimens ordinarily contain some protein
Blood: The test is based on the Pseudo-peroxidase activity of the from moisture and light. When stored in the original container, the Substances that cause abnormal urine color may affect the (<20mg/dL) therefore only persistent elevated levels of urine protein
haem moiety of hemoglobin and myoglobin. The chromogen is product is stable up to the expiry date printed on the label and (or) readability of test pads in urinalysis reagent strips. indicate kidney or urinary tract disease. The persistent results of
oxidized by a hydroperoxide in the presence of haem and changes vial box. trace level or over indicate significance proteinuria and thus further
color from yellow (or greenish yellow) to blue. Replace the bottle cap immediately and tightly after removing Blood:. Elevated specific gravity or protein in urine may reduce the
reactivity of the blood test portion. Microbial peroxidase associated clinical testing is needed to evaluate the significant of results.
Ingredients: diisopropylbenzene dihydro peroxide 26.0%W/W test strips, and keep the vial tightly closed between tests. Do not
tetramethylbenzidine 1.5%W/W remove desiccant from bottle. Do not touch test areas of urine with urinary tract infection may cause false positive results. Ascorbic Nitrite: Normally no nitrite is detectable in urine.
reagent strips. Do not open container until ready to use. acid concentrations (>30 mg/dl) may cause false negatives at the
Bilirubin: Azo-coupling reaction of bilirubin with a diazonium salt low level of blood. Leukocyte: Normally no leukocytes are detectable in urine.
Discoloration or darkening of the test pads may indicate
in an acid medium to form an azodye. Color changes from light tan pH: Urine values generally range from pH 5 to 9.
deterioration. If this is evident, or if test results are questionable Bilirubin: Metabolites of drugs, such as pyridum and selenium,
to beige or light pink.
or inconsistent with expected finding, confirm that the product is which give a color at low pH, may cause false positives. Indican Specific Gravity (SG): The normal SG of urine ranges from 1.001
Ingredients: 2,4-dichloroaniline diazonium salt 0.6%W/W
within its expiration date and is reacting properly using known (indoxyl sulfate) can produce a yellow-orange to red color to 1.035.
Urobilinogen: The test is based on the Ehrlich’s reaction. negative and positive control materials. Do not use after the expiry response, which may interfere with the interpretation of negative
Color changes from light orange-pink to dark pink. date. Note once the canister has been opened, the remaining strips or positive bilirubin readings. Ascorbic acid (> 30mg/dl) may cause
remain stable for up to 6 months.
Ingredients: fast blue B salt 0.2%W/W false negative result.
Performance characteristics are based on clinical and analytical
Ketone: Legal’s test-nitroprusside reaction. Acetoacetic acid in Urobilinogen: The absence of urobilinogen in the specimen cannot studies and depend upon several factors: the variability of colour
an alkaline medium reacts with nitroferricanide to produce a color
Specimen Collection and Preparation perception; the persence or absence of inhibitory and matrix factors
be determined. The test area will react with interfering substances
Collect urine in a clean, dry container that allows complete immersion
change from beige to purple. known to react with Ehrlich’s reagent, such as p-aminosalicylic acid. typically found in urine; and the laboratory conditions in which the
of all the fields on the test strip. Do not add preservatives. Test the
Ingredients: sodium nitroprusside 5.7%W/W Drugs containing azo gantrisin may give a masking golden color. product is used(e.g., lighting, temperature,and humidity). Each
specimen as soon as possible, with the sample well mixed but not
The test is not reliable method for the detection of porphobilinogen. colour block represents a range of values. Because of specimen
Glucose: Glucose oxidase catalyzes the oxidation of glucose to centrifuged. The use of fresh morning urine is recommended for
and reading variability, specimens with analyte concentrations that
form hydrogen peroxide. The hydrogen peroxide thus formed optimal nitrite tests, as well as for the valid determination of bilirubin Ketone: Positive results (trace or less) may occur with highly fall between normal levels may give results at either level. Results
then oxidizes a chromogen on the reaction pad by the action of and urobilinogen, since these compounds are unstable when pigmented urine specimens or those containing large amounts will usually be within one level of the true concentration. The
peroxidase. exposed to light. If immediate testing is not possible, the sample of levodopa metabolites. Some high SG and low pH urine may following list shows the generally detectable levels of the analytes
Ingredients: glucose oxidase (microbial.123U) 1.7%W/W should be stored in the refrigerator, but not frozen, and then brought give false positive result. Phenosulfonphthalein may cause false in contrived urines; however, because of the inherent variability of
peroxidase(horseradish.203U) 0.2%W/W to room temperature before used in the test. Unpreserved urine positive result. clinical urines, lesser concentrations may be detected under certain
tetramethylbenzidine 0.1%W/W at room temperature may undergo pH changes due to microbial
Glucose: High SG (>1.020) with high pH urine and ascorbic acid conditions.
proliferation, which may interfere with protein determination. If
Protein: Protein “error of indicators.” When pH is held constant (more than 40mg/dl) may cause false negative result at the low
cleanly voided specimens are not collected from females, positive
by a buffer, indicator dyes release H+ ions because of the protein
results for leukocytes may be found due to contamination from
level of glucose. Test Pad and Sensitivity (Specificity)
present and change color from yellow (or greenish yellow) to blue- Blood: 10 RBC/µl (0.03mg/dL hemoglobin, Intact
outside the urinary tract. Skin cleansers containing chlorhexidine Protein: False positive results may be found in strongly basic urine
green. RBC)
may affect protein test results if specimen contamination occurs. (pH 9). The interpretation of results is also difficult in turbid urine
Ingredients: tetrabromophenol blue 0.1%W/W Bilirubin: 1 mg/dL (Bilirubin)
specimens.
Urobilinogen: 2 EU/dL (Urobilinogen)
Nitrite: The test is based on the diazotization reaction of nitrite with Instructions for use Nitrite: Ascorbic acid (>30mg/dL) may cause false negative result Ketone: 5-15mg/dL (Acetoacetic acid)
an aromatic amine to produce a diazonium salt. It is followed by The procedure must be followed exactly to achieve reliable results.
with low level of nitrite containing (<0.03mg) urine. The negative Glucose: 50-100mg/dL (Glucose)
an azo-coupling reaction of this diazonium salt with an aromatic Do not compare strips with color chart before the strip is dipped in
result does not always mean that the patient is free from bacteriuria. Protein: 15-30mg/dL (albumin)
compound on the reaction pad. The azo dye produced causes a urine.
Pink spots or pink edges should not be interpreted as a positive Nitrite: 0.05mg/dL (Nitrite ion)
color change form white to pink. 1. Dip the strip into the urine up to the test area for no more than
result. Negative result may occur when urinary tract infections are Leukocytes: 20-25 WBC/µl (Intact and lysed WBCs)
Ingredients: p-arsanilicacid-N-(1-Naphthol)-ethylenediamine two second.
caused by organism which do not contain nitrate reductase; when
1.3%W/W 2. Draw the edge of the strip along the brim of the vessel to
urine has not been retained in the bladder long enough (four hours References
tetrahydroquinoline 0.9%W/W remove excess urine; at this time, don’t make the test areas
or more) for reduction of nitrate to nitrite occur; or when dietary NCCLS ( National Committee for Clinical Laboratory Standard)
touched to the brim of the vessel.Turn the strip on its side
Leukocyte: This test pad contains an indoxyl ester and diazonium nitrate is absent. GP 16-A/ ROUTINE URINALYSIS AND COLLECTION
and tap once on a piece of absorbent material to remove any
salt. It is followed by an azo-coupling reaction of the aromatic TRANSPORATION AND PRESEVATION OF URINE SPECIMENS;
remaining urine; Excessive urine on the strip may cause the Leukocyte: The test result may not always be consistent with
amine formed by leukocytes esterase with a diazonium salt on the TRNTATIVE GUIDELINE VOL 12-NO 26, EC.1992
interaction of chemicals between adjacent reagent pads, so the leukocyte cell number by the microscopic examination. High
reaction pad. The azo dye produced causes a color change from
that an incorrect result may occur. concentration of glucose, high specific gravity, high level of
beige to violet.
3. Compare the colours of the reagent pads exactly after 60 albumin, high concentration of formaldehyde or presence of blood
Ingredients: indoxyl ester 4.3%W/W
seconds (Leukocytes after 90~120 seconds) with the color may cause decreased test results. High concentration of oxalic acid ORDERING INFORMATION
phenyl diazonium salt 0.4%W/W
chart on the vial label under good light. While comparing, keep of trace of oxidizing agents may cause false negative results.
pH: Double indicator system. Indicator’s methyl red and the strip horizontally to prevent possible mixing of chemicals CATALOG NO. QUANTITY
pH: If the excessive urine is remain on the strip because of
bromothymol blue are used to give distinct color changes from when excessive urine is present. 1100 100 100 Test Strip
improper test procedure, it is possible that the acidic buffer in
orange to green to blue. (pH 5.0 to 9.0)
protein portion comes out and affect the pH portion, then pH result
Ingredients: methyl red 3.3%W/W
may be decreased than the actual. This phenomenon is called ”run-
bromthymol blue 55.0%W/W
over effect.”
308 309

Spectrum Diagnostics Products List 2014-2015

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Product Inserts 2014

Egyptian Company For Biotechnology (S.A.E.)

Corrosive (C) Calculation Newborns:

Batch Code/Lot number CAUTION. Consult instructions Others

LOT Batch Code/Lot number CAUTION. Consult instructions

Reagents also contain non-reactive stabilizers and Expected Values

REAGENT PREPARATION AND STORAGE Expected Values Men Women

to lower the risk of heart disease and coronary artery disease. A

HDL + CHOD + CHER PVS/PEGME

(Single Reagent) EC REP Authorised Representative

Assay Principle Notes

Reagents 25 Tests 50 Tests 100 Tests

B. Glycosylated hemoglobin (GHb) Separation

C. Total Hemoglobin (THb) fraction

Important Note: QUALITY CONTROL

Run to run (Reproducibility)

Drabkin’s Solution EC REP Authorised Representative

Drabkin’s Solution EC REP Authorised Representative Use by/Expiration Date Precision

Intended Use Reagent Preparation Storage and Stability

Reagent Preparation DA = A2 - A1 For Sample and Standard

Precautions and Warnings Mean (µmol/L) 45.54 171.62

GLDH 8.0 K U/L Zero adjustment Against Air Haemolysis

Peroxidase >3000 U/L

REF Catalogue Number for use

METHOD / REACTION PRINCIPLE: Stability of Working reagents

α-Naphthol + FRTR-salt Azo dye PROCEDURE:

REAGENTS: (Concentrations in the test) I-Manual assay for Total ACP

Reagent A 1.0 ml (Pre-heated At 37°C)

The sealed reagents are stable up to the indicated expiry date

Linearity 750 U/L

Consult instructions for use Manufactured by n 20 20

MgCl2 2.0 mmol/L Working 1.0 ml ( or add 0.5 ml R1 + 0.5 ml R2 ) Lipemia

Serum and Plasma Icterus

(Single Reagent) - GALG2-CNP EC REP Authorised Representative

Goods Buffer pH 6,0 50 mmol/L Sample 3 72,1 1,700 0,934

Storage and Stability

E.C.2.6.1.1. LOT Batch Code/Lot number

Working 1 ml ( or add 0.5 ml R1 + 0.5 ml R2 ) Expected value (at 37 oC)

Reagent (R) Urine 24 h urine 95 – 240 mmol/24h

REF: 228 001 50 Tests

Components ( concentrations in the test ) Procedure

Reagent 1 (buffer pH 4.5) SD 2.1 1.9 2.12 1.36

Mix, and incubate for 5 to 10 minutes at 20 – 25 C. Read the

Lipemia 4. Viollier MA, Gschwind H, Schläpfer P. Neue

Specimen collection and preparation

REAGENTS PREPARATION AND STABILITY For Urine

Interfering Substances BIBLIOGRAPHY

Standard Potassium 5.00 mmol/L

Reagent Storage and Stability Linearity

LOT Batch Code/Lot number CAUTION. Consult instructions Waste Disposal

Precautions and Warnings Expected Values

System Parameters Spectrum Diagnostics does not interpret the results of a

Standard (St.) 200 mg/dl (30.6 mmol/l) Quality Control

For further information, refer to the Aspartate aminotransferase Linearity

Precautions and Warnings Expected values

REF: 326 001 (1 vial ) 1 x 5 ml

Description Expected Values

Procedure 3. Richtlinien der Bundesärztekammer zur Qualitätssicherung in

REF: 327 001 (1 vial ) 1 x 5 ml

Procedure 3. Richtlinien der Bundesärztekammer zur Qualitätssicherung in

REF: 328 001 (1 vial ) 1 x 5 ml

REF: 356 001 1 X 0.5 ml

Preparation Refer to the package inserts of the reagent kits respectively