Requirements
- Personal protective equipment
- Lab coat
- Protective Apron
- Lab-grade gloves
- Shoe covers
- Protective face shield or protective goggles
- Spill kit
- Workstation
- Clean & disinfected workstation
- Tube discarder box
- Sharps discarder
- Calibrated Pipettes
- Calibrated water-bath
- Coagulation calibrated electrical centrifuge
- Thermometer
- Optimally functioning digital stopwatch
- Labelling marker
- Light source (Bright lamp)
- PT result recording sheet
- Calculator
- Test related
- Multiple clean glass tubes
- Citrated Control Platelet Poor Plasma (C-PPP)
- Citrated Test/Patient's Platelet Poor Plasma (T-PPP)
- QC verified Thromboplastin Reagent
Procedure
- Step 1
- Label a clean glass test tube as Thromboplastin
- Add >200 μL Thromboplastin reagent in this tube
- Place this test tube in water-bath at 37 0C
- Step 2
- Label 4 clean glass test tubes as following:
- C1 → Citrated Control PPP
- C2 → Citrated Duplicate Control PPP
- T1 → Citrated Test PPP
- T2 → Citrated Duplicate Test PPP
- Step 3
- Take C1 labelled tube & add 100 μL Citrated Platelet Poor Plasma (PPP) into it
- Incubate C1 labelled tube in water-bath at 370C for 02 mins
- Step 4
- Take out 200 μL 370C pre-incubated Thromboplastin reagent (from Thromboplastin Labelled Tube) and add it to C1 labelled tube (which has completed 2 mins incubation in water-bath in Step 3)
- Immediately start the Digital Stopwatch
- Step 5
- From 9 seconds onwards remove the C1 labelled tube from water-bath to see for clot formation at regular intervals (every 2 seconds)
- Note the time when the clot has formed & stop the Stopwatch
- Step 6
- Repeat the procedure as per above steps in C2 labelled tube
- Take the average of the readings from C1 & C2 labelled tubes
- Step 7
- Now, repeat all the above mentioned steps in duplicate with Test PPP from the patient in T1 & T2 labelled tubes
- Record the average of the readings from T1 & T2 labelled tubes
Reporting of Results
- Results are reported as per following two formats:
- The mean (average) of the PT readings run in duplicate in
seconds
- PT ratio between Citrated Test & Control PPP
- For PT Value of Citrated Control PPP in Routine
- GMNPT is calculated as the mean of PT
for 20 healthy individuals as under:
- 10 samples from healthy males
- 10 samples from healthy females
- No sample from those receiving any oral anticoagulant
- GMNPT is calculated for every new Thromboplastin reagent
- The mean (average) of the PT readings run in duplicate in seconds
- PT ratio between Citrated Test & Control PPP
- GMNPT is calculated as the mean of PT for 20 healthy individuals as under:
- 10 samples from healthy males
- 10 samples from healthy females
- No sample from those receiving any oral anticoagulant
- GMNPT is calculated for every new Thromboplastin reagent
Interpretation of Results
- Causes of Isolated PT prolongation
with normal APTT (Click For Details)
- FVII deficiency
- Start of oral anticoagulants
(Warfarin)
- Lupus anticoagulant: Some
thromboplastins are sensitive to LA
- Mild liver impairment or vitamin K deficiency
- Depending upon reagent used mild
deficiency of II, V, X
- Causes of Isolated Shortening of PT
- Following treatment with rFVIIa
(NovoSeven or SevenFact)
- FVII deficiency
- Start of oral anticoagulants (Warfarin)
- Lupus anticoagulant: Some thromboplastins are sensitive to LA
- Mild liver impairment or vitamin K deficiency
- Depending upon reagent used mild deficiency of II, V, X
- Following treatment with rFVIIa (NovoSeven or SevenFact)
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