SOP-Advia Centaur XP

R. K.
LIFE SERVICES PRIVATE LIMITED

(Department of Laboratory Medicine)
STANDARD OPERATING
PROCEDURE
(Biochemistry ADVIA
CENTAUR XP)
COPY: MASTER COPY
HOLDER: QUALITY MANAGER
ISSUE NO: 04
ISSUE DATE: 09.03.2020

R. K. LIFE SERVICES PRIVATE LIMITED
Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY
AMENDMENT SHEET
Sl. Page Section/ Date of Amendment Reasons Signature Signature

No. No. Clause Amendment Made of
No. of QM Laboratory
Director
01 All All 09.03.2020 Issue No 03 Change in
has been Analyser
withdrawn
and Issue No
04 has been
issued in the
system
02 03-36 Reportable 18.08.2022 Reportable Adequacy
Interval Interval has Raised in
changed Assessment
Issue No. 04 Issue Date: 09.03.2020 Prepared By: Copy No. Page 1 of 1
Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I
TABLE OF CONTENTS
LAST
CHAPTER SL. REVISION
TITLE REVISION PAGE No.
No. No. NO
MADE
- 1. TABLE OF CONTENTS 00 00 01 - 02
2. TSH 00 00 03 - 05
3. T3 00 00 06 - 08
4. T4 00 00 09 - 11
5. FT4 00 00 12 - 14
6. PSA 00 00 15 - 17
I 7. PROLACTIN 00 00 18 - 20
8. TOTAL IgE 00 00 21 – 23
9. FERRITIN 00 00 24 - 26
10. FT3 00 00 27 – 29
11. VITAMIN D 00 00 30 – 33
12. VITAMIN B 12 00 00 34 - 36
Machine Operation
Procedure: ACESS
II 13. 00 00 01
IMMUNOASSAY
SYSTEM
QUALITY CONTROL
III 14. 00 00 04
PROCEDURE

LAST
CHAPTER SL. REVISION
TITLE REVISION PAGE No.
No. No. NO
MADE
CALIBRATION
IV 15. 00 00 01
PROCEDURE
SAFETY AND
PRECAUTIONS
V 16. 00 00 01
PROCEDURE

NAME OF TEST: T.S.H.

a. PURPOSE OF THE EXAMINATION:
For in vitro diagnostic use in the quantitative determination of thyroid-
stimulating hormone (TSH, thyrotropin) in serum using the ADVIA
Centaur, ADVIA Centaur XP, and ADVIA Centaur XPT systems.
b. PRINCIPLE AND METHOD OF THE PROCEDURE USED FOR

EXAMINATION:
The ADVIA Centaur® TSH assay is a two-site sandwich immunoassay
using direct chemiluminometric technology, which uses constant
amounts of two antibodies. The first antibody, in the Lite Reagent, is a
monoclonal mouse anti-TSH antibody labeled with acridinium ester. The
second antibody, in the Solid Phase, is a polyclonal sheep anti-TSH
antibody which is covalently coupled to paramagnetic particles.
c. PERFORMANCE CHARACTERISTICS:
0.05 μIU/mL to 150 µIU/ml
Method: CHEMILUMINOMETRIC TECHNOLOGY
d. TYPE OF SAMPLE:
Serum
Storage: can be stored at 2 to 80C for 24-hours
e. PATIENT PREPARATION: QMSP-15

f. TYPE OF CONTAINER AND ADDITIVES:
Plain evacuated tube or Microtainer, No anticoagulant.
g. REQUIRED EQUIPMENT AND REAGENTS:

1. Auto Immunoassay Analyser (ADVIA CENTAUR XP) 2. Centrifuge 3.
Test Tube Racks 4. Sample Container 5. Variable Micro Pipettes 0-100
and 100 – 1000 µl 6. Micro tips 7. Sample cup 0.5 ml 8. Sample cup 2.0
ml 9. TSH Kit

h. ENVIRONMENTAL AND SAFETY CONTROLS

1. Temp: 200C-250C 5. Centrifuge
2. Humidity: not more than 80%-90 % 6. Stability of Power Supply
3. Dust free.
4. Wrong sampling.
Safety precaution
Details safety controls in chapter – V
i. CALIBRATION PROCEDURE:
Calibrated with TSH Calibrator i.e.. Recalibration is done if, (i) L-J Curve
shows nonconformance (ii) EQAS levels show nonconformance. (iii)
Change of reagent lot. (iv) When calibration curve expired.
All instruments are calibrated as per NABL 112 requirements. It is
described in Chapter-IV.
j. PROCEDURAL STEPS:
Procedure of collection of samples, processing, transport and storage
prior to test is described in QMSP-15 (Procedure for sample handling).
Operation of instrument is mentioned in Chapter – II
k. QUALITY CONTROL PROCEDURES:

For internal quality assessment Biorad Control two levels are run. Also
participate in BIORAD EQAS.
Detail Quality Control Procedure is mentioned in Chapter -III
l. INTERFERENCES AND CROSS REACTION:
m. PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS

INCLUDING, WHERE RELEVANT, THE MEASUREMENT
UNCERTAINTY OF MEASURED QUANTITY VALUES:
Instrument automatically calculates the result. CV% is calculated after
plotting the control points monthly in the L.J charts. MU% is calculated
as CV% X 1.96.

n. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION

VALUES:
Age Limit Male Female
1 – 11 months 0.8 -6.3 (μIU/mL) 0.8 -6.3 (μIU/mL)
1 – 05 years 0.7 – 5.9 (μIU/mL) 0.7 – 6.0 (μIU/mL)
Adults 0.34 – 5.6 (μIU/mL) 0.34 – 5.6 (μIU/mL)
o. REPORTABLE INTERVAL OF EXAMINATION RESULTS:

0.010–150 μIU/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs
q. INSTRUCTIONS FOR DETERMINING QUANTITATIVE RESULTS

WHEN A RESULT IS NOT WITHIN THE MEASUREMENT INTERVAL:
Sample having TSH value more than 150 µIU/ml should be manually
diluted and re-run.
r. ALERT/CRITICAL VALUES:
Not Applicable
s. LABORATORY CLINICAL INTERPRETATION:

Thyrotropin or thyroid-stimulating hormone is a glycoprotein with a
molecular weight of 28,000 to 30,000 daltons.
In cases of hyperthyroidism, TSH level is severely inhibited and may even
be undetectable.
In cases of primary hypothyroidism, TSH levels are always much higher
than normal and thyroid hormone levels are low.
t. POTENTIAL SOURCES OF VARIATION:

a) Patient on treatment
b) High dose hook effect (>150 µIU/ml).
c) Critically ill patient. In such patient TSH assay as a routine
must not be done.
d) Deterioration of reagents.
u. REFERENCE: Kit literature

NAME OF TEST: T3
For in vitro diagnostic use in the quantitative determination of
triiodothyronine (T3) in serum using the ADVIA Centaur, ADVIA Centaur
XP, and ADVIA Centaur XPT systems. Measurements of triiodothyronine
are used in the diagnosis and treatment of thyroid disease.

EXAMINATION:
The ADVIA Centaur® T3 assay is a competitive immunoassay using direct
chemiluminescent technology. T3 in the patient sample competes with a
T3 analog, which is covalently coupled to paramagnetic particles in the
Solid Phase for a limited amount of acridinium ester-labeled monoclonal
mouse anti-T3 antibody in the Lite Reagent.
0.1 to 8.0 ng/mL
d. TYPE OF SAMPLE:
Serum
Storage: Can be stored at 2 to 80C for 24-hours


ml 9. T3 Kit

h. ENVIRONMENTAL AND SAFETY CONTROLS:

3. Absolutely dust free.
4. Wrong sampling.
Safety precaution
Calibrated with T3 Calibrator i.e.. Recalibration is done if, (i) L-J Curve
Operation of instrument is mentioned in Chapter – II.


as CV% X 1.96.


VALUES:
1 – 11 months 1.0 – 2.17 (ng/mL) 0.98 – 2.20 (ng/mL)
1 – 05 years 1.0 – 2.12 (ng/mL) 0.92 – 2.14 (ng/mL)
6 – 10 years 0.9 – 2.03 (ng/mL) 1.0 – 2.0 (ng/mL)
11 – 15 years 0.8 – 1.95 (ng/mL) 0.70 – 1.90 (ng/mL)
16 – 20 years 0.72 – 1.85 (ng/mL)) 0.60 – 1.80 (ng/mL)
Adults 0.87 – 1.78 (ng/mL) 0.87 – 1.78 (ng/mL)

0.1–8 ng/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having T3 value more than 8.0 ng/ml should be manually
diluted and re-run.
Not Applicable

Hypo/Hyper Thyroidism / Euthyroidism / Sick euthyroid syndrome

b) High dose hook effect; if it is more than 8 ng/ml.
c) Critically ill patient. In such patient T3 assay as a routine must
not be done.
u. REFERENCE: Kit literature (Beckman system)

NAME OF TEST: T4
For in vitro diagnostic use in the quantitative determination of thyroxine
(T4) in serum using the ADVIA Centaur, ADVIA Centaur XP, and ADVIA
Centaur XPT systems. Measurements of thyroxine are used in the
diagnosis and treatment of thyroid disease.

EXAMINATION:
The ADVIA Centaur® T4 assay is a competitive immunoassay using direct
chemiluminescent technology. T4 in the patient sample competes with T 4,
which is covalently coupled to paramagnetic particles in the Solid Phase,
for a limited amount of acridinium ester-labeled monoclonal mouse anti-
T4 antibody in the Lite Reagent.
0.3 μg/dL to 30.0 μg/dL
d. TYPE OF SAMPLE:
Serum
Storage: Can be stored at 2 to 80C for 24-hours


ml 9. T4 Kit


4. Wrong sampling.
Safety precaution:
Calibrated with T4 Calibrator i.e.. Recalibration is done if, (i) L-J Curve


as CV% X 1.96.


VALUES:
1 – 11 months 1.0 – 2.17 (ng/mL) 0.98 – 2.20 (ng/mL)
1 – 05 years 1.0 – 2.12 (ng/mL) 0.92 – 2.14 (ng/mL)
6 – 10 years 0.9 – 2.03 (ng/mL) 1.0 – 2.0 (ng/mL)
11 – 15 years 0.8 – 1.95 (ng/mL) 0.70 – 1.90 (ng/mL)
16 – 20 years 0.72 – 1.85 (ng/mL)) 0.60 – 1.80 (ng/mL)
Adults 0.87 – 1.78 (ng/mL) 0.87 – 1.78 (ng/mL)

0.3–30 μg/dL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having T4 value more than 30.0 µg/dl should be manually
diluted and re-run.
Not Applicable
Hypo/Hyper Thyroidism

b) High dose hook effect, if the value is > 30 µl/dl
c) Critically ill patient. In such patient T4 assay as a routine must
not be done.

NAME OF TEST: FREE T4

For in vitro diagnostic use in the quantitative determination of free
thyroxine (FT4) in serum or plasma (heparinized or EDTA), using the
ADVIA Centaur, ADVIA Centaur XP, and ADVIA Centaur XPT systems.
Measurements of free thyroxine are used in the diagnosis and treatment
of thyroid disease.

EXAMINATION:
The ADVIA Centaur® FT4 assay is a competitive immunoassay using
direct chemiluminescent technology. FT4 in the patient sample competes
with acridinium ester-labeled T4 in the Lite Reagent for a limited amount
of biotinylated polyclonal rabbit anti-T4 antibody.
Biotin-labeled anti-T4 is bound to avidin that is covalently coupled to
paramagnetic particles in the Solid Phase.
0.1 ng/dL to 12.0 ng/dL
d. TYPE OF SAMPLE:
Serum
Storage:
Can be stored at 2 to 80C for 24-hours
e. PATIENT PREPARATION: QMSP 15
f. TYPE OF CONTAINER & ADDITIVES:

 Plain evacuated tube or Microtainer, No anticoagulant.

1.Auto Immunoassay Analyser (ACCESS) 2. Centrifuge 3. Test Tube
Racks 4. Sample Container 5. Variable Micro Pipettes 0-100 and 100 –
1000 µl 6. Micro tips 7. Sample cup 0.5 ml 8. Sample cup 2.0 ml 9. FT4
Kit


4. Wrong sampling.
Safety precaution
Calibrated with FT4 Calibrator i.e.. Recalibration is done if, (i) L-J Curve
k. QUALITY CONTROL PROCEDURE:

Detail Quality Control Procedure is mentioned in Chapter –III
l. INTERFERENCES and cross REACTION:

as CV% X 1.96.

n. BIOLOGICAL REFERENCE INTERVALS

1 – 25 days 0.76 – 1.80 (ng/dL) 0.85 – 1.73 (ng/dL)
1 – 24 months 0.68 – 1.26 (ng/dL) 0.70 – 1.25 (ng/dL)
9 – 10 years 0.75 – 1.10 (ng/dL) 0.75 – 1.13 (ng/dL)
11 – 14 years 0.65 – 1.01 (ng/dL) 0.68 – 1.05 (ng/dL)
Adults 0.74 – 1.17 (ng/dL) 0.68 – 1.06 (ng/dL)

0.1–12 ng/dL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs
WHEN A RESULT IS NOT WITHIN THE MEASUREMENT
INTERVAL;
Sample having FT4 value more than 12.0 ng/dl should be manually
diluted and re-run.
Not Applicable
Hypo/Hyper Thyroidism / Euthyroidism / Sick euthyroid syndrome.

c)
Critically ill patient. In such patient FT4 assay as a routine must not
be done.
d)
Deterioration of reagents.

NAME OF TEST: PSA

This in vitro diagnostic assay is intended to quantitatively measure
prostate-specific antigen (PSA) in human serum using the ADVIA
Centaur®, ADVIA Centaur XP, and ADVIA Centaur XPT systems. This
assay is indicated for the measurement of serum PSA in conjunction with
Digital Rectal Exam (DRE) as an aid in the detection of prostate cancer in
men aged 50 years and older. This assay is further indicated as an aid in
the management (monitoring) of patients with prostate cancer.

EXAMINATION:
The ADVIA Centaur PSA assay is a two-site sandwich immunoassay using direct
chemiluminometric technology, which uses constant amounts of two antibodies. The first
antibody, in the Lite Reagent, is a polyclonal goat anti-PSA antibody labeled with
acridinium ester. The second antibody, in the Solid Phase, is a monoclonal mouse anti-
PSA antibody, which is covalently coupled to paramagnetic particles.
0.01 ng/dL to 100.0 ng/mL
d. TYPE OF SAMPLE:
Serum
Storage:


1000 µl 6. Micro tips 7. Sample cup 0.5 ml 8. Sample cup 2.0 ml 9. PSA
Kit


4. Wrong sampling.
Safety precaution
Calibrated with PSA Calibrator i.e.. Recalibration is done if, (i) L-J
Curve shows nonconformance (ii) EQAS levels show nonconformance.
(iii) Change of reagent lot. (iv) When calibration curve expired.


Serum specimens that are . . . Demonstrate ≤ 5% change in results up to . . .
Hemolyzed 500 mg/dL of hemoglobin
Lipemic 1000 mg/dL of triglycerides
Icteric 40 mg/dL of bilirubin

as CV% X 1.96.
0.27 – 3.42 ng/ml


0.01–25 ng/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs
INTERVAL;
Sample having PSA value more than 100.0 ng/ml should be manually
diluted and re-run.
Not Applicable
PSA is a glycoprotein which belongs to the kallikrein family.
PSA level rise in prostatic pathologies such as being prostatic hyperplasia
(BPH) or prostrate cancer. Testing for PSA and its evolution is useful for
monitoring and controlling the efficacy of prostatic carcinoma therapy.
Determination of PSA levels enables the detection of the onset of
metastases or the persistence of disease following prostrate cancer
therapy. Am elevated PSA level after therapy or a persistently high level
during therapy indicates residual or recurrent disease.

c) Critically ill patient. In such patient PSA assay as a routine must not
be done.

NAME OF TEST: PROLACTIN

For in vitro diagnostic use in the quantitative determination of prolactin
in serum using the ADVIA Centaur, ADVIA Centaur XP, and ADVIA
Centaur XPT systems.

EXAMINATION:
The ADVIA Centaur® Prolactin assay is a two-site sandwich
immunoassay using direct chemiluminometric technology, which uses
constant amounts of two antibodies. The first antibody, in the Lite
Reagent, is a polyclonal goat anti-prolactin antibody labeled with
acridinium ester. The second antibody, in the Solid Phase, is a
monoclonal mouse anti-prolactin antibody, which is covalently coupled to
paramagnetic particles.
0.3 ng/mL to 200.0 ng/mL
d. TYPE OF SAMPLE:
Serum
Storage:


1.Auto Immunoassay Analyser (ADVIA CENTAUR XP) 2. Centrifuge 3.
ml 9. Prolactin Kit


4. Wrong sampling.
Safety precaution
Calibrated with Prolactin Calibrator i.e.. Recalibration is done if, (i) L-J
Curve shows nonconformance (ii) EQAS levels show nonconformance.
(iii) Change of reagent lot. (iv) When calibration curve expired.


Serum specimens that are . . . Have an insignificant effect on the assay up to . . .
Hemolyzed 500 mg/dL of hemoglobin
Lipemic 1000 mg/dL of triglycerides
Icteric 20 mg/dL of bilirubin

as CV% X 1.96.


Male: 2.64 – 13.13 ng/ml
Female:
Pre Menopause: 3.34 – 26.72 ng/ml
Post Menopause: 2.74 – 19.64 ng/ml
0.3–200 ng/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs
INTERVAL;
Sample having Prolactin value more than 200.0 ng/mL should be
manually diluted and re-run.
Not Applicable
Prolactin is a glycoprotein with a molecular weight of approximately
23,000 daltons.
Many factors control the secretion of prolactin. TRH (thyrotropin-
releasing-hormone) and VIP (vasocative intestinal peptide) stimulate
prolactin secretion.
Exogenous factors such as physical exercise, stress, diet and
hypoglycemia can cause an increase in prolactin levels.

c) Critically ill patient. In such patient Prolactin assay as a routine
must not be done.

NAME OF TEST: Total IgE

For in vitro diagnostic use in the quantitative determination of total IgE
in serum using the ADVIA Centaur, ADVIA Centaur XP, and ADVIA
Centaur XPT systems.

EXAMINATION:
The ADVIA Centaur® Total IgE assay is a two-site sandwich
immunoassay using direct chemiluminometric technology, which uses
constant amounts of two antibodies to IgE. The first antibody, in the Lite
Reagent, is a goat anti-human IgE antibody labeled with acridinium
ester. The second antibody, in the Solid Phase, is a mouse anti-human
IgE antibody, which is covalently coupled to paramagnetic particles.
1.5 IU/mL to 3000.0 IU/mL
d. TYPE OF SAMPLE:
Serum
Storage:


1000 µl 6. Micro tips 7. Sample cup 0.5 ml 8. Sample cup 2.0 ml 9.
Total IgE Kit


4. Wrong sampling.
Safety precaution
Calibrated with IgE Calibrator i.e.. Recalibration is done if, (i) L-J Curve


as CV% X 1.96.


0 – 150 IU/ml
1.5–3000 IU/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs
INTERVAL;
Sample having IgE value more than 3000.0 IU/ml should be manually
diluted and re-run.
Not Applicable
IgE have been associated with type I immediate hypersensitivity. Atopic
individuals can produce fairly high quantities of IgE after exposure to
specific allergens.

c) Critically ill patient. In such patient IgE assay as a routine must not
be done.

NAME OF TEST: FERRITIN

For in vitro diagnostic use in the quantitative determination of ferritin in
serum or plasma using the ADVIA Centaur ®, ADVIA Centaur XP, and
ADVIA Centaur XPT systems to aid in the diagnosis of iron deficiency
anemia and iron overload.

EXAMINATION:
The ADVIA Centaur Ferritin assay is a two-site sandwich immunoassay using direct
chemiluminometric technology, which uses constant amounts of two anti-ferritin
antibodies. The first antibody, in the Lite Reagent, is a polyclonal goat anti-ferritin
antibody labeled with acridinium ester. The second antibody, in the Solid Phase, is a
monoclonal mouse anti-ferritin antibody, which is covalently coupled to paramagnetic
particles.
0.5 ng/mL to 1650 ng/mL
d. TYPE OF SAMPLE:
Serum


ml 9. Ferritin Kit


3. Dust free.
4. Wrong sampling.
Safety precaution
Calibrated with Ferritin Calibrator i.e.. Recalibration is done if, (i) L-J
Curve shows nonconformance (ii) EQAS levels show nonconformance. (iii)


as CV% X 1.96.


VALUES:
23.9 – 336.2 ng/ml
0.5–1650 ng/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having Ferritin value more than 1650 ng/ml should be manually
diluted and re-run.
Not Applicable

The drop in serum ferritin levels can indicate an iron deficiency prior to
the appearance of anemia.
The level of serum ferritin acts as an indicator of the quantities or iron in
the human body. It also closely correlated to the level of iron in the bone
marrow.

b) Critically ill patient. In such patient Ferritin assay as a
routine must not be done.
c) Deterioration of reagents.

NAME OF TEST: FT3

For in vitro diagnostic use in the quantitative determination of free
triiodothyronine (FT3) in serum using the ADVIA Centaur, ADVIA Centaur
XP, and ADVIA Centaur XPT systems. Measurements of free
triiodothyronine are used in the diagnosis and treatment of thyroid
disease.

EXAMINATION:
The ADVIA Centaur® FT3 assay is a competitive immunoassay using
direct chemiluminescent technology. FT3 in the sample competes with a
T3 analog, which is covalently coupled to paramagnetic particles in the
Solid Phase for a limited amount of a combination of acridinium ester-
labeled monoclonal mouse anti-T3 antibodies in the Lite Reagent.
0.2 pg/mL to 20 pg/mL
d. TYPE OF SAMPLE:
Serum


ml 9. FT3 Kit

3. Dust free.

4. Wrong sampling.
Safety precaution
Calibrated with FT3 Calibrator i.e.. Recalibration is done if, (i) L-J Curve


as CV% X 1.96.

VALUES:
2.5 – 3.9 pg/ml


0.2–20 pg/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having FT3 value more than 20 pg/mL should be manually
diluted and re-run.
Not Applicable

FT3 plays an important diagnostic role in the following cases:
 Hyperthyroidism: Basedow’s disease or toxic adenoma
 Monitoring of patients with hyperthyroidism treated with
Thyroxine anf antithroid agents.
 Low-T3 syndrome.

b) Critically ill patient. In such patient FT3 assay as a routine
must not be done.

NAME OF TEST: Vitamin D

The ADVIA Centaur® Vitamin D Total (VitD) assay is for in vitro diagnostic
use in the quantitative determination of total 25(OH)vitamin D in human
serum and plasma (EDTA, lithium-heparin, sodium-heparin) using the
ADVIA Centaur, ADVIA Centaur XP, and ADVIA Centaur XPT systems.
The ADVIA Centaur VitD assay is intended as an aid in the determination
of vitamin D sufficiency.

EXAMINATION:
The ADVIA Centaur VitD assay is an 18-minute antibody competitive
immunoassay that uses an anti-fluorescein monoclonal mouse antibody
covalently bound to paramagnetic particles (PMP), an anti-25(OH)vitamin
D monoclonal mouse antibody labeled with acridinium ester (AE), and a
vitamin D analog labeled with fluorescein.
An inverse relationship exists between the amount of vitamin D present
in the patient sample and the amount of relative light units (RLU)
detected by the system.
4.2 ng/mL to 150 ng/mL
d. TYPE OF SAMPLE:
Serum


ml 9. Vit-D Kit


3. Dust free.
4. Wrong sampling.
Safety precaution
Calibrated with Vitamin D Calibrator i.e.. Recalibration is done if, (i) L-J



as CV% X 1.96.

VALUES:
Deficiency - < 20 ng/ml
Insufficiency – 20 – 30 ng/ml
Sufficiency – 30 – 70 ng/ml
Non toxic overdose – 70 – 150 ng/ml

4.2–150 ng/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having Vitamin D value more than 150 ng/mL should be
Not Applicable

Vitamin D is a steroid hormone involved in the intestinal absorption of
calcium and the regulation of calcium homeostasis.
Physiologically Vitamin D levels result not only from dietary uptake but
also can be produced from a cholesterol precursor. 7-dehydrocholesterol,
in the skin during sun exposure.
The concentration of Vitamin D decreases with age and a deficiency is
common among elderly persons.
Clinically applications of 25 (OH) Vitamin D measurements are in the
diagnosis and therapy control of post menopausal, Osteoporosis,
Pregnancy, neonatal hypocalcemia and hyperparathyroidism.
Vitamin D intoxicans mostly occurs during a large intake of
pharmaceutical preparations of Vitamin D and may lead to
Hypercalcemia, hypercalcuria and nephocalcinosis in susceptible infants.


b) Critically ill patient. In such patient Vitamin D assay as a

NAME OF TEST: Vitamin B12

For in vitro diagnostic use in the quantitative determination of vitamin
B12 in serum or plasma using the ADVIA Centaur ®, ADVIA Centaur XP,
and ADVIA Centaur XPT systems.

EXAMINATION:
The ADVIA Centaur VB12 assay is a competitive immunoassay using
direct chemiluminescent technology in which vitamin B12 from the
patient sample competes with vitamin B12 labeled with acridinium ester
in the Lite Reagent, for a limited amount of purified intrinsic factor,
which is covalently coupled to paramagnetic particles in the Solid Phase.
The assay uses Releasing Agent (sodium hydroxide) and DTT to release
the vitamin B12 from the endogenous binding proteins in the sample and
cobinamide to prevent rebinding after the Solid Phase is added to the
sample.
45.0 pg/mL to 2000 pg/mL
d. TYPE OF SAMPLE:
Serum


1. Auto Immunoassay Analyser (ACCESS-2) 2. Centrifuge 3. Test Tube
1000 µl 6. Micro tips 7. Sample cup 0.5 ml 8. Sample cup 2.0 ml 9. Vit-
B12 Kit


3. Dust free.
4. Wrong sampling.
Safety precaution
Calibrated with Vitamin B12 Calibrator i.e.. Recalibration is done if, (i) L-J


as CV% X 1.96.


VALUES:
180 – 914 pg/ml

45–2000 pg/mL
p. TAT
Regular: 8 hrs
Emergency: 2 hrs

Sample having Vitamin B12 value more than 2000 pg/mL should be
Not Applicable

Vitamin B12 is the name given to any one of a group of substances termed
cobalamines. It is a co-enzyme that is involved in two very important
metabolic functions vital to normal cells growth and DNA synthesis.
Cobalamin deficiency leads to Megaloblastic anaemia and demyelination
of large nerve fibres of spinal cord. Sources of vitamin B 12 are liver,
eggs, meat, fish shelfish and other diary products.
Decreased level:
 Lack of intrinsic factor: gastrectomy, atrophic gastritis.
 Malababsorption as in regional ileitis, resected bowel, tropical
sprue, cellac disease.
 In dietary deficiency in vegetarian.

b) Critically ill patient. In such patient Vitamin B12 assay as a

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY CHAPTER - II
MACHINE OPERATING PROCEDURE

MACHINE OPERATING PROCEDURE: ADVIA CENTAUR XP ANALYSER
Main menu
[F1]Test Request/Progress
Test Request
/Progress
Enter Tray ID, Then [Enter]
Next sample Position

Enter Sample ID, Then [Enter]
Type optional Patient ID, then [Enter]
Select test for Test list by typing
Test ID More tests
Press[+]
All samplesNo
Entered? [Enter]
Y
[F1]Get/Load Tray to load samples
[F1]Done when finished

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY CHAPTER - III
QUALITY CONTROL PROCEDURE:

Purpose:
Quality control (QC) is concerned with the analytical phase of QA. The purpose
of this document is to monitor the overall reliability and monitoring the validity
of laboratory results.
Scope:
All immunoassay tests under the scope of Accreditation
RESPONSIBILITY: Consultant Pathologist
Following Procedure is followed in Biochemistry.

A. Regular use of QC material of renowned QC kit manufacturer
B. Participation in EQAS / inter-laboratory comparison
C. Retesting of retained sample
D. Correlations of results
A. Regular use of QC material of renowned QC kit manufacturer:

Laboratory runs QC as per Quality Control Plan (RKLS/FM/70) i.e. daily one level of control for all
the parameters under scope. Laboratory interprets QC results by using statistical technique such
as L-J chart. Where any parameter value is found to deviate Westgard Multi QC rules, cause is
identified & corrective action is taken. Consultant reviews it and records are maintained
accordingly.
Calculation of CV %: The standard deviation (SD) of a set of results divided by the mean result is
expressed as a percentage CV. In automated analyzer percentage CV is auto calculated along with
L-J chart.
GENERAL PROCEDURE AND TERMINOLOGY:

Running commercial control:
Advia Centaur XP
Two levels control are run once daily for all of the parameters
Rev. No.: 00 Rev. Date: Nil Approved By: Issued By

PROCEDURE FOR HANDLING INTERNAL COMMERCIAL CONTROL:
 Receive the sample and confirm Lot No and Expiry Date with reference values specific to
the machine.
 Make a note of the storage temperature.
 Controls are stored at 2-8oC in specified Biochemistry Refrigerator
 Vials are kept at room temperature for a maximum period of 1 hour while running tests.
 Record sample validity period after opening or reconstitution.
 Prepare aliquots (if applicable) and store in recommended temperature.
 Internal Controls are run on a daily basis immediately after the specific machines are switched
on and in perfect running mode. Only authorized personnel in each department run controls.
 Values observed are recorded and plotted in L-J charts.
 Observe for any violation of Multi QC Deviation Rule and proceed accordingly.
INTERPRETATION OF QUALITY CONTROL DATA
QC Results are reviewed & if any of the parameters run out of 2SD or do not follow any of the MULTI
QC RULES described below, then a QC Level is run for the second time. If the results still do not fall
within the expected range, then the test parameter is calibrated, QC checked & samples run. All the
QC values are checked and filed and reviewed by the Consultant Pathologist.

The rules to follow when 1 level QC material are used:

Reject QC if:
13S: One control observation exceeding the mean ± 3s; primarily sensitive to random error.
22S: Two consecutive control observation exceeding the same mean plus 2 s or mean minus 2 s
limit; sensitive to systematic error.
10X: 10 consecutive control observations falling on one side of the mean (above or below, with no
other requirement on size of the deviations); sensitive to systematic error.
41s: Four consecutive values on either side of mean are within 1SD to 2SD (41S).
The rules to follow when 2 level QC material are used:

Reject QC if:
1.
Either QC value is outside 3 SD (1 3 S)
2.
Both QC value are outside 2 SD on the same side, but within 3 SD (2 2 S)
3.
Different between both QC value is > 4 SD i.e. one level QC is > 2 SD and other level QC is
<2SD (R 4 S)
4.
Ten consecutive value of the same level QC are >/< the mean, but within 2 SD (10x)
5.
Five consecutive value of one level QC and five consecutive value of another level QC are
>/< the mean but within 2 SD (10x).
6.
Four consecutive values on either side of mean are within 1SD to 2SD
Procedure to be followed when QC results are “out of control” (> ± 2 SD).
FOLLOW STEP WISE TILL QC RESULTS ARE “IN CONTROL”

1. Repeat with same aliquot of QC.
2. Repeat with new aliquot of QC.
3. Reconstitute new QC.
4. Calibrate with current calibration pack and same reagent.
5. Calibrate with new calibration pack and same reagent.
6. Check generation no. of slide/ lot no. Cartridge/ reagent expiry / on board reagent expiry.
7. Use new reagent.
8. Calibrate with new calibration pack and new reagent
9. Contact respective Application Specialist.

B. Participation in EQAS / Inter-Laboratory Comparison:

The laboratory participates in EQAS with Bio-Rad. Before testing of EQAS sample daily QC results
are reviewed, if results are found satisfactory then EQAS sample is tested. ILC is done with NABL
accredited labs once in a month.
C. Retesting of retained sample:

Retesting of the sample is done daily. Primary sample is chosen at a random and tested within 24
hrs. Consultant Pathologist reviews retesting results and record is maintained.
D. Correlations of results:
Different Tests when results indicate the same pathology are used to confirm test results when
necessary. Both inter and intradepartmental correlation is done. Pathologist reviews the record

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY CHAPTER - IV
CALIBRATION PROCEDURE:
ADVIA CENTAUR XP IMMUNOASSAY AUTOANALYSER
 Machine calibrated by standardization procedure.

 Machine is calibrated twice in a year or as per the manufacturer’s requirement.
 All parameter are calibrated with the calibrator provided in the kit as and when required
Rev. No.: 00 Rev. Date: Nil Approved By: Issued By:

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY CHAPTER - V
SAFETY AND PRECAUTIONS PROCEDURE:
 Safety instructions of different instruments and chemicals given by manufacturers are followed strictly.
 General safety precautions of biohazard are observed.
 Samples are handled with utmost care.
 Samples are handled wearing gloves, facemask and apron.
 Reagents and chemicals are kept according to the company instructions
Issue No. 04 Issue Date: 09.03.2020 Issued By: QM Copy No. Page 1
Rev. No.: 00 Rev. Date: Nil Approved By: LD Prepared By: QM

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R. K.

LIFE SERVICES PRIVATE LIMITED

COPY: MASTER COPY

HOLDER: QUALITY MANAGER

ISSUE DATE: 09.03.2020

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY

Sl. Page Section/ Date of Amendment Reasons Signature Signature

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

NAME OF TEST: T.S.H.

b. PRINCIPLE AND METHOD OF THE PROCEDURE USED FOR

Method: CHEMILUMINOMETRIC TECHNOLOGY

e. PATIENT PREPARATION: QMSP-15

g. REQUIRED EQUIPMENT AND REAGENTS:

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

h. ENVIRONMENTAL AND SAFETY CONTROLS

k. QUALITY CONTROL PROCEDURES:

l. INTERFERENCES AND CROSS REACTION:

m. PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

n. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION

o. REPORTABLE INTERVAL OF EXAMINATION RESULTS:

q. INSTRUCTIONS FOR DETERMINING QUANTITATIVE RESULTS

s. LABORATORY CLINICAL INTERPRETATION:

t. POTENTIAL SOURCES OF VARIATION:

u. REFERENCE: Kit literature

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

b. PRINCIPLE AND METHOD OF THE PROCEDURE USED FOR

Method: CHEMILUMINOMETRIC TECHNOLOGY

e. PATIENT PREPARATION: QMSP-15

f. TYPE OF CONTAINER AND ADDITIVES:

g. REQUIRED EQUIPMENT AND REAGENTS:

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

h. ENVIRONMENTAL AND SAFETY CONTROLS:

k. QUALITY CONTROL PROCEDURES:

l. INTERFERENCES AND CROSS REACTION:

m. PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

n. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION

o. REPORTABLE INTERVAL OF EXAMINATION RESULTS:

q. INSTRUCTIONS FOR DETERMINING QUANTITATIVE RESULTS

s. LABORATORY CLINICAL INTERPRETATION:

t. POTENTIAL SOURCES OF VARIATION:

u. REFERENCE: Kit literature (Beckman system)

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

b. PRINCIPLE AND METHOD OF THE PROCEDURE USED FOR

Method: CHEMILUMINOMETRIC TECHNOLOGY

e. PATIENT PREPARATION: QMSP-15

f. TYPE OF CONTAINER AND ADDITIVES:

g. REQUIRED EQUIPMENT AND REAGENTS:

Rev. No.: 01 Rev. Date: 18.08.2022 Approved By: Issued By

Doc No. RKLS/SOP/01 SOP BIOCHEMISTRY Chapter - I

h. ENVIRONMENTAL AND SAFETY CONTROLS

k. QUALITY CONTROL PROCEDURES: