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    Coombs Test

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    unilabreynosa
    The document describes the Indirect and Direct Coombs tests, which are used to detect antibodies in a patient's serum or attached to red blood cells, respectively. The Indirect Coombs test looks for antibodies in the serum that could cause hemolytic disease of the newborn, while the Direct Coombs test detects antibodies attached to red blood cells that could indicate a transfusion reaction, drug sensitization, or autoimmune hemolytic anemia. The procedures involve adding patient samples to control or test cells, incubating, washing cells, adding anti-human serum, and examining for agglutination to identify the presence of antibodies.

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    Coombs Test

    Uploaded by

    unilabreynosa
    467 views11 pages
    The document describes the Indirect and Direct Coombs tests, which are used to detect antibodies in a patient's serum or attached to red blood cells, respectively. The Indirect Coombs test looks for antibodies in the serum that could cause hemolytic disease of the newborn, while the Direct Coombs test detects antibodies attached to red blood cells that could indicate a transfusion reaction, drug sensitization, or autoimmune hemolytic anemia. The procedures involve adding patient samples to control or test cells, incubating, washing cells, adding anti-human serum, and examining for agglutination to identify the presence of antibodies.

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    21642794-Coombs-Test

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    The document describes the Indirect and Direct Coombs tests, which are used to detect antibodies in a patient's serum or attached to red blood cells, respectively. The Indirect Coombs test looks for antibodies in the serum that could cause hemolytic disease of the newborn, while the Direct Coombs test detects antibodies attached to red blood cells that could indicate a transfusion reaction, drug sensitization, or autoimmune hemolytic anemia. The procedures involve adding patient samples to control or test cells, incubating, washing cells, adding anti-human serum, and examining for agglutination to identify the presence of antibodies.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    467 views11 pages

    Coombs Test

    Uploaded by

    unilabreynosa
    The document describes the Indirect and Direct Coombs tests, which are used to detect antibodies in a patient's serum or attached to red blood cells, respectively. The Indirect Coombs test looks for antibodies in the serum that could cause hemolytic disease of the newborn, while the Direct Coombs test detects antibodies attached to red blood cells that could indicate a transfusion reaction, drug sensitization, or autoimmune hemolytic anemia. The procedures involve adding patient samples to control or test cells, incubating, washing cells, adding anti-human serum, and examining for agglutination to identify the presence of antibodies.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
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    of 11

    COOMBS TEST

    Indirect Coombs test (Indirect Antiglobulin test):


     This test is performed to detect presence of Rh-antibodies
    or other antibodies in patients serum in case of the
    following:
    1. To check whether an Rh-negative women (married to Rh-
    positive husband) has developed Anti Rh-antibodies

    2. Anti D may be produced in the blood of any Rh-negative


    person by exposure to D antigen by-
     Transfusion of Rh positive blood
     Pregnancy, if infant is Rh positive (if father is Rh-positive)
     Abortion of Rh-positive fetus.
    Requirements:
     Test tubes: (10x75 mm)
     Pasteur pipettes
     Incubator
     Centrifuge
    Specimen: Serum (need not be fasting)
    Reagents:
    1. Antihuman serum
    2. Anti-D serum
    Additional Requirements:
     Coombs control cells
    A. Make a pooled ‘O’ Rho (D) positive cells from at least three different ‘O’
    positive blood samples.
    B. Wash these cells three times in normal saline (these cells should be
    completely free from serum with no free antibodies). Make 5 % saline
    suspension of these cells
    Procedure:

    1. Label three test tubes as ‘T” (test serum) PC (Positive


    control) and NC (negative control).

    2. In the tube labelled as ‘T’, add two drops of Anti-D serum

    3. In the tube ‘PC’ add one drop of saline

    4. Add one drop of 5 % saline suspension of the pooled ‘O’


    Rho (D) positive cells in each tube.

    5. Incubate all the three tubes for one hour at 37°C


    Procedure: (cont.)

     Wash the cells three times in normal saline to remove


    excess serum with no free antibodies, (in the case of
    inadequate washings of the red cells, negative results
    may be obtained).
     Add two drops of Coombs serum (anti human serum)
    to each tube. Keep for 5 minutes and then centrifuge
    at 1,500 RPM for one minute.
     Resuspend the cells and examine macroscopically as
    well as microscopically
    Test Interpretation:
    Observations Conclusions

    1 Positive Control (A) Agglutination Correctly performed


    (PC) test procedure.
    (B) No Agglutination Coombs serum may
    not be proper.
    Repeat the test again.

    2 Negative It should show no agglutination, since saline does


    control not contain Anti-D or any other antibodies.
    (NC)
    3 Test (Serum) (T) (A) Agglutination (andPatients serum
    if PC results are contains Anti-D.
    correct)

    (B) No Agglutination Patients serum does


    not contain Anti-D.
    Direct Coombs test (direct antiglobulin test):
     This test is performed to detect anti-D antibody or other
    antibodies attached to the red cell surface within the blood
    stream.
     This occurs in the following circumstances:
     When there is a Rh-positive baby in the womb of a sensitized Rh-
    negative women; the antibodies produced in the mothers serum
    cross the placenta and after entering the baby's blood stream,
    these antibodies will attach to the baby's Rh-positive red blood
    cells. These coated (or sensitized) cells are clumped and removed
    from the circulation, causing hemolytic anemia (Hemolytic Disease
    of the Newborn: Erythroblastosis Fetalis). When the baby is born,
    the baby's blood is collected (or cord blood is collected from
    umbilical cord) and tested by the anti globulin Coombs test (direct)
    to detect anti D antibodies coated on red blood cells.
     Transfusion reactions
     Drug induced red cells sensitization
     Autoimmune hemolytic anemia
    Requirements: (same as that for Indirect Coombs test)
     Test tubes: (10x75 mm)
     Pasteur pipettes
     Incubator
     Centrifuge

    Specimen: Blood drawn into EDTA is preferred but oxalateed, or clotted,


    citrated whole blood may be used (specimen need not be fasting
    sample)
    Procedure:
    1. Prepare a 5 % suspension in isotonic saline of the red blood cells to be
    tested.

    2. With clean Pasture pipette add one drop of the prepared cell
    suspension to a small tube.
    3. Wash three times with normal saline to remove all the traces of serum.
    4. Decant completely after the last washing
    5. Add two drops of Antihuman serum.

    6. Mix well and centrifuge for one minute at 1500 RPM.

    7. Resuspend the cells by gentle agitation and examine macroscopically


    and microscopically for agglutination.

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