PENGENDALIAN

SPESIMEN DI
MAKMAL
Noorfaizah binti Ghazali
Pegawai Sains (Kimia Hayat) C41
Unit Patologi & Transfusi
Hospital Selama, Perak
Pre-analytical Factors

Controllable
variables

Non-Controllable
variables
Controllable variables
Patient Complication
Diet, Exercise, Lifestyle,
Drugs, Travel, Posture,
Prolonged bedrest
Specimen Transportation
Diet, Exercise, Lifestyle,
Drugs, Travel, Posture,
Prolonged bedrest

Spesimen collection &

handling
Patient’s detail, test
requested, location, diagnosis,
collection date & time
Non-Controllable variables

Biological Influences Long-term cyclical changes

Heredity, age, sex, race Seasonal influences,
Menstrual cycle, Underlying
medical conditions

Enviromental Factors
Altitute, Ambient
temperature, Residence
SPECIMEN COLLECTION & HANDLING
Information required on laboratory requisition
Patient name
Identification number (I/C no)
Location (wad/clinic/hospital)
Test requested
Diagnosis/diet
Collection date/time
Doctor’s identification/signature
 TYPE OF SPESIMEN
-Blood
-Urine
-Cerebrospinal fluid (CSF)
-Semen
-Peritoneal, pericardial and pleural fluid
-Sweat
-Feces
-Saliva
-Synovial fluid
-Amniotic fluid
-Tissue and organs (including malignant)
 BLOOD : SITE OF BLOOD COLLECTION
• Most frequently • Dangerous • Infants, old
used for all •Requires patient
blood chemistry considerable skills • Small
•For blood gas
amount
Arterial Capillary
Venipuncture
puncture blood
BLOOD : SITE OF BLOOD COLLECTION
 TOURNIQUET & DRAW TECHNIQUE
Placed 3 to 4 inches above venipuncture site
- Too high veins not prominent
- Too close hematoma
< 1 minute,if longer → hemoconcentration
(↑K, Prot,PCV)

Blood collection near site of infusion

→ contamination / dilution
Traumatic draw from vessel wall injury
→ ↑CK, myoglobin, and K
Central venous catheter – Composition may be affected by
infused fluids.
Blood drawn from central venous catheter and peripheral vein
can have different values even if properly collected.
 ORDER OF BLOOD DRAW

Blood culture collected → sodium citrate tube

→plain tube → Heparin tube →EDTA →
Fluoride tube (oxalate/EDTA)
 DRAW VOLUME
Underfilling Specimen → dilution → Erroneous PT,PTT and RBC’S
morphology (shrink)
Overfilling → Inadequate anticoagulation, fibrin formation,
microclots, platelet clumping
 MIXING & INVERSION OF TUBE
Insufficient mixing cause microclots
↓
Affect : platelets, HCT, hemoglobin
and fibrin formation, instrument
probe
 SERUM CLOTTING TIME
Serum should be clotted before centrifugation
(30-60 min)
If inadequate :
Problem for many instruments because of
latent fibrin formation ( probe clogging) and
error result
Serum should separated within 2 hours:
Prolonged contact time of serum or plasma
with RBCs
↓
Exchange of compounds
• Blood specimen should not be transferred form one tube to
another.
• Different tube may contain different types of additive
 Anticoagulants and preservatives for blood
Heparin: as Na, K, Li, NH4+
• Action: accelerate antithrombin III thus prevent fibrin formation
• Could affect T3 and T4 analysis

Ethylenediaminetetraacetic acid (EDTA) : as N2 , K2 or K3 salt

• A chelating agent
• Action: forms complex with Ca salt
• Unsuitable: Ca and iron analysis

Sodium fluoride
• antiglycolytic agent, but a weak anticoagulant (usually come with K oxalate)
• Action: inhibit enzymes in glycolysis
• Affects urea nitrogen analysis
• Alternative replacement: Na iodoacetate (does not affect urease)

Sodium citrate
• action: Chelates calcium
• affects phosphate, Ca
measurement
 GEL/PLAIN TUBE
-Routine Chemistry (RP/LFT/FLP/CE/UA)
- IMMUNOASSAY
- (Hormone/Tumor Marker)
- - IRON STUDY

EDTA TUBE
-Whole Blood Sample
- FBC/ RETIC COUNT/HBA1C/IPTH/AMMONIA/LACTATE

SODIUM CITRATE TUBE

-Coagulation Sample
(PT/APTT)

FLOURIDE TUBE
-GLUCOSE (Fasting/Random/2HPP)
 Moderately Specimen Hemolysed
hemolysed

Factors that contribute to hemolysis:

Normal serum
 Forcing the blood through needle/
Grossly
hemolysed
unsuitable needle size
 Shaking the tube or bulb too vigorously
after blood collection
 Centrifuging blood samples at high speed
before completion of clotting
 Freezing or thawing of blood
Slightly
 Using unclean tubes with residual
hemolysed detergent
 Presence of water in the container
 Putting tourniquet too tight
 Alcohol were not completely dry
 Too much pressure being applied during
blood taking or transferring blood into
tubes
 Specimen Lipaemic

Should be avoided
Interfere with spectrophotometric, turbidic
measurements→false result
Will showed high TG value
Can be avoided by:
- fasting (9-12 hours) before blood were taken
- ultracentrifugation

NORMAL LIPAEMIC
 URINE COLLECTION
Type of urine specimens:
Random specimen (untimed)
Timed specimen - Pre-determined interval of time –
1, 4, or 24 hours
“Clean-catch specimen” – bact. culture
Catheter specimen – Microbiological examination in
critically ill, taken from bladder through urethra
Suprapubic tap – taken directly form bladder, using
22G spinal needle

*Genitalia cleaned to avoid chances of

contamination,
especially for urine culture.
 URINE PRESERVATIVES
 Glacialacetic acid – Aldosterone, catecholamines, cortisol,
estrogens, metanephrine.
 Boric acid – Homovanillic acid, hCG, VMA
 Concentrated HCl – Amino acids, calcium, copper,
catecholamines, 5HIAA, VMA, hydroxyproline, magnesium,
mercury, metanephrines, oxalate, nitrogen
 Mild base – Porphyrins, amino levulinic acid
 Chloroform – Organic acids
 Nitric acid - Mercury
SPECIMEN TRANSPORTATION
Timing
-Some specimens must be
transported immediately
after collection. Ex. Arterial
blood gas to minimize O2
consumption
-Specimen must centrifuged
and separated within 2 hours
-Excessive transport time to
analysis result in hemolysis
Temperature
-Appropriate temperature
Chilling ( 2 to 8 oC). not
recommended for whole
blood beyond 2 hours.
-Chilling specimen required
for catecolamin, ammonia,
lactate, PTH and gastrin
-Some samples need to be
protected from light as
bilirubin
-Transport in leak proof
plastic bags
 Put all the samples
belongs to same patient
in a single BIOHAZARD
plastic bag before sent
to lab
-ASSUME THAT ALL THE
SAMPLES ARE
BIOHAZARD!
BLOOD CULTURE
 Paeds/ O2/ AnO2

 Keep the blood culture

bottle at RT until it
reaches the lab
 Sent the blood culture
bottle at any time to
the lab (24 hrs) Adult Pediatric

Adults : 8-10 ml
DO NOT LABEL ON
THE BARCODE!!! Paeds : 1-3 ml
BLOOD CULTURE
Can I draw blood from catheter?

-Whenever possible draw blood from a peripheral vein

and NOT from catheter except in catheter related
bacteremia (CRBSI), need to withdraw from both
periphery vein and from a catheter (within one hour)

-Use aseptic technique

-Correct amount:adult,8-10mls,paeds:1-3mls
 No specimen specimen Container Volume storage after notes
collection
1. Blood C/S Adult (2 Bottle) Adult Room temp. in wards Not Do not press the skin after
1. Bactec plus + aerobic (blue) 3-10ml in each >48 hr disinfection unless with
2. Bactec plus anaerobic(gold) bottle gloves
Paediatric (1 bottle) paeds 2. Surface of the bactec
1. Paeds Bactec plus + (pink) 1.0-3.0ml bottle must wipe with 70%
of alcohol swab before the
blood insert
2. Urine C/S Sterile urine container/bottle - Refrigerator 1.Collect early morning
2°C - 8°C specimen
2. place the specimen in
ice if the potter system
delayed
3. CSF C/S Bijou sterile bottle 1 - 2 ml Room temp. in wad Send the specimen as
soon as possible

4. Body Fluids C/S Sterile Container/bottle 3-4 ml Room temp in wad Send the specimen as
soon as possible

5. Stool C/S a. Cary-Blair media Groundnut size Room temp Early morning stool
(small amount) preferred or
Rectal swab
Handling of MerscoV sample
MerscoV Request Form
How to collect the
sample??
Transportation of sample – Triple packaging
REJECTION
STUDY
 Requested Form Specimen
 No/wrong specimen
 No request form  No/wrong labelled
 Wrong request form REJECTION CRITERIA  Leaking
 No/wrong patient’s  Hemolysed
name  Clott
 No/wrong patient’s ID  Test not offered
 Ward/Clinic was not  Insufficient
written
Others  Wrong container
 Contaminate
 No diagnosis/clinical
 Specimen not suitable to be  The way of handling sample
history did not meet the
 No request test was analysed
 Specimen sent at wrong time requirement
written  No transport to sent out the  Poor smear
 Time & date specimen specimen
taken was not written  Wrong blood group
 No signature/stamping  Wrong R/N
of requested doctor  Specimen sent repeatedly
 machine breaks down
INCOMPLETE REQUEST FORM
 Clottedsample
Clotted sample
Factors that contribute to blood clotting
i) Improper mixing of the tubes
ii) Inadequate volume of the blood – inaccurate
ratio of anticoagulant & blood

Overfilling Underfilling
 Contaminated sample
 Transferring blood sample
from one tube to another
(different tube contains
different
preservative/anticoagulant)

 Follow the order of draw to

prevent contimation od the
samples
KEPENTINGAN KEPUTUSAN UJIAN MAKMAL

 membantu anggota klinikal dalam

membuat diagnosis terhadap pesakit

 Untuk tujuan pengurusan pesakit

 Untuk tujuan penyaringan sesetengah

penyakit
“GOOD IN, GOOD OUT”
GOOD QUALITY OF SAMPLES WILL GIVE A GOOD (ACCURATE) RESULT