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    5 Hema 2 Laboratory Manual-PT

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    Janielle Fajardo

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    5 Hema 2 Laboratory Manual-PT

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    75 views4 pages

    5 Hema 2 Laboratory Manual-PT

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    Janielle Fajardo

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    MEDT 20: HEMATOLOGY 2 LABORATORY

    Prothrombin Time

    Learning Outcomes

    After the completion of this module, you will be able to:


    1. Demonstrate the proper way of preparing blood samples for coagulation
    studies
    2. Explain the significance of prolonged prothrombin time

    Introduction

    Prothrombin Time (PT) measures the time where the clot is formed, this is a
    coagulation study test commonly used for screening for extrinsic factor deficiency,
    monitoring oral anticoagulant therapy and quantitative determination of the extrinsic
    coagulation factors deficiency. This test was described by Quick in 1935 and the test was
    initially often referred to as 'Quick's Prothrombin Time.' Prothrombin is one of many the
    coagulation factors that plays a crucial role for clot formation.

    Principle

    Tissue thromboplastin, in the presence of calcium ions


    and Factor VII, activates the extrinsic pathway of coagulation.
    When a mixture of tissue thromboplastin and calcium ions is
    added to normal anticoagulant plasma, the clotting mechanism
    is initiated and a clot will form within a specified time period. If a
    deficiency exists within the extrinsic pathway, the time required
    for clot formation will be prolonged. The degree of prolongation
    is proportional to the severity of single factor deficiency, or in a
    cumulative deficiency of all the factors involved.

    Materials

    1. Citrated Evacuated tube


    2. Centrifuge
    3. Serological pipette (0.1 and 0.2 ml)
    4. Wassermann tube (2)

    JMF 2024
    MEDT 20: HEMATOLOGY 2 LABORATORY

    5. 37 degree Celsius

    6. Thromboplastin reagent
    7. Tissue paper
    8. Timer

    Preparation of specimen for PT:

    1. Collect venous blood using citrated tube.


    2. Centrifuge at 1500 rpm for five minutes.
    3. Separate the plasma by placing it into a clean dry tube.

    HEMOSTAT PROTHROMBIN TIME

    Principle:
    The on-stage PT measures the clotting time of plasma after adding a source of tissue factor
    (Thromboplastin) and calcium. The recalcification of plasma in the presence of tissue factor
    generate activated factor X, with the consequent formation of thrombin and ultimately a fibrin
    clot.

    Reagent:

    Thromboplastin reagent (stored at 2-8 C)


    -Lyophilized extract of rabbit brain
    -Calcium chloride

    Reagent preparation:

    Reconstitute with exactly 2 ml distilled water. Agitate gently until the solution is complete.
    After reconstitution, the reagent is stable for seven days when stored at 2-8 C, 24 hours at
    15 – 25 C and 24 hours at 37 C. Storing the reagent between 2-8 C when it is not in use is
    recommended.

    Test Procedure
    1. Perform samples and controls in duplicates
    2. Prewarm the Hemostat thromboplastin SI reagent at 37 C
    3. Pipette 0.1 mL of plasma/Control into a prewarmed test tube
    4. Incubate at 3-5 minutes at 37 C
    5. Add PT-SI reagent
    6. Start timer with the addition of reagent
    7. Record the time required for clot formation

    JMF 2024
    MEDT 20: HEMATOLOGY 2 LABORATORY

    Normal Value: 10-14 seconds

    Date
    Name/s

    Year/Section/Cluster

    5 points

    1. Give at least 5 conditions wherein PT is prolonged?

    2. Why is the prothrombin time employed in monitoring patients who is under

    warfarin therapy?

    JMF 2024
    MEDT 20: HEMATOLOGY 2 LABORATORY

    References:

     Sadang M.M and Llanera, F.R (2015) Laboratory Manual in Hematology


     Elaine M. Keohane, Larry J. Smith, Jeanine M. Walenga 2016 Rodak's
    Hematology Clinical Principles and Applications 5th Edition
     Screening tests in Haemostasis: The Prothrombin Time [PT]. (n.d.).

    JMF 2024

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