PROTHROMBIN TIME

• Prothrombin time is the plasma clotting time obtained when excessive thromboplastin and
optimum calcium are added to citrated plasma under standardized conditions
• PT is essentially a test of the extrinsic pathway of clotting and common pathway
• It defects deficiencies in factor I, II, V, VII and X
• The test is least sensitive to factor II and insensitive to factor IX and to other factors involved in
the intrinsic thromboplastin generation
• PT is frequently used to monitor oral anticoagulant
• Prolonged PT is also seen:
o Vitamin K deficiency, certain liver diseases, specific deficiencies, DIC, presence of
circulation anticoagulants and fibrin/fibrinogen split products
• Sodium citrate is the preferred anticoagulant
• The test should be performed within two hours after blood collection

REAGENT PREPARATION

• Reconstitute with 2 mL distilled water

• Agitate gently until the solution is complete
• After reconstitution, the reagent is stable for seven days when stored at 2 to 8 C, 24 hours at 15
to 25 C and 24 hours at 37C
• Storing the reagent between 2 to 8 C when it is not in use is recommended

TEST PROCEDURE

• Perform samples and controls in duplicates

• Prewarm the HemoStat Thromboplastin-SI reagent at 37C
• Pipette 0.1 mL of plasma/control into a prewarmed test tube
• Incubate at 3 to 5 minutes at 37 C
• Add PT-SI reagent
• Start timer with the addition of reagent
• Record the time required for clot formation
• NV: 10-14 seconds

ACTIVATED PARTIAL THROMBOPLASTIN TIME

• aPTT is a routine test for screening coagulation disorders in the intrinsic pathway by measuring
factors VIII, IX, XI, XII, and also Prekallikrein (PK) or fletcher factor but not platelets and factor
XIII
• APTT is also used to detect the presence of circulating anticoagulant or inhibitors, and to
monitor heparin therapy
• Sodium citrate is the preferred anticoagulant
• The test should be performed within 2 hours after blood collection
 Materials

• Two way needle

• Citrated evacuated tube
• Centrifuge
• 0.3mL serological pipette
• 0.4mL serological pipette
• Tissue paper
• Wasserman tube
• 37C water bath
• APTT reagent
• Calcium chloride
• Timer

Preparation of specimen for APTT

1. Collect venous blood in a citrated tube
2. Centrifuge at 1,500 rpm for 5 minutes
3. Separate the plasma by placing it into a clean dry tube

HEMOSTAT ACTIVATED PARTIAL THROMBOPLASTIN TIME

• Determination of aPTT utilizing Ellagic Acid Activator

• The aPTT is a simple and versatile test which is sensitivev to deficiencies of all plasma clotting
factors except factor VII
• However, it is principally usedto detectdeficiencies in stage 1of the coagulation mechanism,
namely factors VIII, IX, XI and Prekallikrein (or Fletcher factor)

Test Principle
• The HemoStat aPTT-EL test is performed by adding aPTT reagent containing a plasma activator
and phospholipids to the test specimen; the phospholipids serve as a substitute for platelets
• This mixture is incubated for three minutes 37C for optimum activation
• The incubated mixture is then racalcified with a calcium chloride solution and clot formation is
timed. The aPTt-el reagent can also be used to perform quantitative factor assays.

Content and Reagenet Preparation

• HemoSTat aPTT-EL
o Chloroform extract of rabbit brain
o Ellagic acid
• CaCl2
o Calcium chloride (0.02 mol/L)

Test Procedure
1. Perform samples and controls in duplicates
2. Prewarm the HemoStat CaCl2 (0.02 M) to 37 C
3. Pipetteinto 0.1 mL of plasma/control prewarmed test tube
4. Incubate for 1 to. 2 minutes at 37 C
5. Add 0.1 mL aPTT-EL reagent
6. Incubate for 3 minutes at 37C
7. Add 0.1 mL of prewarmed CaCl2
8. Start timer with addition of CaCl2
9. Record time required for clot formation
10. Calculate the mean time of duplicate aPTT determination for each test plasma and report to the
nearest. 0.1 seconds
• Normal Reference range: 23.4 to 36.2 seconds