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    Direct Coombs

    Uploaded by

    sanjibbhunia408

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Direct Coombs

    Uploaded by

    sanjibbhunia408
    6 views2 pages

    Original Title

    DIRECT COOMBS

    Copyright

    © © All Rights Reserved

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    PDF, TXT or read online from Scribd

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    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    6 views2 pages

    Direct Coombs

    Uploaded by

    sanjibbhunia408

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    DIRECT COOMBS, INDIRECT COOMBS AND CHECK CELL PREPARATION

    CHECK CELL PREPARATION

    **Material Required :-

    * Diluted Anti D (Monoclonal IGG Antibodies) (1:64 Ratio).


    * O Positive Pooled RBC 5 % Suspension.
    * Preservatives and Modified Alsever’s Solution.

    ** 3 Test Tube Label :-


    1. O Positive Pooled RBC 5 % Suspension.
    2. Diluted Anti D ( 1 : 64).
    3. Check Cell ( Coombs Control Cell).

    ** 5 % Sample Suspension O Positive :-


    1 Part Cell Wash Blood : 19 Part Normal Saline = 20

    ** Diluted Solution of Anti D (Monoclonal IgG Only) :-


    1:64 Ratio = 20 ul Anti D + 1260 ul Normal Saline. = 64

    ** Step :-
    Put 1 Drop of 5 % RBC Suspension in Tube Laballed as Check Cell + After That Put 1 Drop of
    Diluted Anti D Solution. Mix Well And Incubation at 37 0 C For 45 Minutes. After 3 Time Cell
    Wash For Normal Saline.

    ** Quality Control For Check Cell:-


    Check Cell 1 Drop and Coombs Reagent 1 Drop Mix Well .
    Agglutination Clumps to Valid Check Cell.
    No Agglutination Invalid Check Cell.
    DIRECT COOMBS TEST

    ** Step:- Now Do Cell Wash of RBC 3 Times With Normal Saline.

    ** EDTA Blood :- 100 ul / Normal Saline 400 ul.

    ** 5 % Sample Suspension :- 1 Part Cell Wash Blood + 19 Part Normal Saline = 20

    ** Step :- 1 Drop of 5 % Sample Suspension + 2 Drop of Coombs Reagent Mix Well and
    Centrifuge at 1000 RPM to 1 Minutes.

    ** Add 1 Drop Check Cell to Coombs Negative Test Tube Agglutination Clumps Appear Valid
    Test. Or No Agglutination No Clumps invalid Test.

    INDIRECT COOMBS TEST

    **1. Plasma / Serum of Sample.


    2. Antihuman Globulin Reagent.
    3. Check Cell.

    ** Take Blood Sample and Centrifuge 3000 RPM for 4 Minutes. Separate Plasma / Serum to
    Another Tube (Patient / Sample Serum).

    ** Prepare and Take 5 % Cell Suspension of O Positive (+ve) RBC.


    ** 3 Test Tube Label :-
    1. Test (Sample Plasma + O Positive(+ve) Suspension).
    2. Positive Control ( Anti D + O Positive(+ve) Suspension).
    3. Negative Control (Saline + Positive(+ve) Suspension).
    ** Step :-
    Add 2 Drop Plasma Sample in Test Tube.
    Add 2 Drop of Anti D In Positive Control Tube.
    Add 2 Drop of Normal Saline in Negative Control Tube.
    Add 1 Drop 5 % Cell Suspension of O Positive (+ve) Cell to Each Tube.
    Mix Well and Incubation at 370 C for 30 Minutes. After Give Normal Saline Wash 3 Times of
    RBC. Discard the Supernatant by Pipette.
    Add 2 Drop of Coombs Reagent to Each Tube and Mix Well all the Tube. And Incubation
    Again for 5 Minutes at 370 C. After Incubation Again Centrifuge for 1 Minutes at 1000 RPM.
    After Mix Well Microscope for Agglutination Clumps it Present Positive Test or No
    Agglutination Clumps it Negative Test.

    ** Add 1 Drop Check Cell to Coombs Negative Test Tube Agglutination Clumps Appear Valid
    Test. Or No Agglutination No Clumps invalid Test.

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