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    9BBK Lec9 - Coombs Test MQA 2019-09-26 08 - 15 - 27

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    gothai sivapragasam
    The document discusses the Coombs test, which is used to detect antibodies bound to red blood cells. It defines the direct and indirect Coombs tests. The direct test detects antibodies bound to red blood cells in vivo, and is used to diagnose immune hemolytic anemia. The indirect test detects antibodies in serum that could bind to red blood cells in vitro, and is used for blood typing and screening for antibodies in pregnant women. Both tests use anti-human globulin to cause agglutination of antibody-coated red blood cells.

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    9BBK Lec9 - Coombs Test MQA 2019-09-26 08 - 15 - 27

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    gothai sivapragasam
    34 views18 pages
    The document discusses the Coombs test, which is used to detect antibodies bound to red blood cells. It defines the direct and indirect Coombs tests. The direct test detects antibodies bound to red blood cells in vivo, and is used to diagnose immune hemolytic anemia. The indirect test detects antibodies in serum that could bind to red blood cells in vitro, and is used for blood typing and screening for antibodies in pregnant women. Both tests use anti-human globulin to cause agglutination of antibody-coated red blood cells.

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    9BBK Lec9- Coombs test MQA 2019-09-26 08_15_27

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    The document discusses the Coombs test, which is used to detect antibodies bound to red blood cells. It defines the direct and indirect Coombs tests. The direct test detects antibodies bound to red blood cells in vivo, and is used to diagnose immune hemolytic anemia. The indirect test detects antibodies in serum that could bind to red blood cells in vitro, and is used for blood typing and screening for antibodies in pregnant women. Both tests use anti-human globulin to cause agglutination of antibody-coated red blood cells.

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    Download as pptx, pdf, or txt
    34 views18 pages

    9BBK Lec9 - Coombs Test MQA 2019-09-26 08 - 15 - 27

    Uploaded by

    gothai sivapragasam
    The document discusses the Coombs test, which is used to detect antibodies bound to red blood cells. It defines the direct and indirect Coombs tests. The direct test detects antibodies bound to red blood cells in vivo, and is used to diagnose immune hemolytic anemia. The indirect test detects antibodies in serum that could bind to red blood cells in vitro, and is used for blood typing and screening for antibodies in pregnant women. Both tests use anti-human globulin to cause agglutination of antibody-coated red blood cells.

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    © All Rights Reserved
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    Download as pptx, pdf, or txt
    of 18

    BACHELOR OF BIOMEDICAL SCIENCES

    BASIC TRANSFUSION SCIENCE AND BLOOD BANKING


    BMC 125

    CHAPTER 9: ANTIGLOBULIN TESTING (COOMB’S )

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Learning Outcome

    At the end of the lecture, students able to

    • Define the Coombs test


    • Direct Coombs test
    • Indirect Coombs test
    • Mechanism and clinical use of coombs test

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    • Antiglobulin test (AGT)
    • Anti human globulin test (AHGT)
    two clinical blood test used :
    ~ immunohematology
    ~ immunology
    • 1st described in 1945:Robin Coombs
    • two type Coombs tests:
    >>direct Coombs test (DCT)/direct antiglobulin test (DAT)
    Detection of sensitization of RBCs in vivo
    >>indirect Coombs test (ICT)/indirect antiglobulin test
    (IAT)
    Detection of sensitization of RBCs in vitro

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    • In certain diseases or conditions an
    individual's blood may contain IgG antibodies
    that can specifically bind to antigens on the
    red blood cell (RBC) surface membrane.
    • Their circulating red blood cells (RBCs) can
    become coated with IgG alloantibodies and/or
    IgG autoantibodies.
    • Complement proteins (C3d and C3b) may
    subsequently bind to the bound antibodies.

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Mechanism
    The two Coombs tests are based on the fact that:
    I. Anti-human antibodies, which are produced by immunizing
    non-human species with human serum, will bind to human
    antibodies, commonly IgG or IgM.
    II. Animal anti-human antibodies will also bind to human
    antibodies that may be fixed onto antigens on the surface of
    red blood cells(RBCs)
     The appropriate test tube conditions can lead to agglutination
    of RBCs . The phenomenon of agglutination of RBCs is important
    because the resulting clumping of RBCs can be visualised
    ~ when clumping is seen the test is positive
    ~ when clumping is not seen the test is negative

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Direct Coombs test (DCT)
    Direct antiglobulin test
    ~ used to detect if antibodies or complement system factors
    have bound to RBC surface antigens in vivo.
    ~ The DAT is not currently required for pre-transfusion testing
    but may be included by some laboratories

    Examples of diseases that give a positive direct Coombs test

    The direct Coombs test is used clinically when immune-mediated


    hemolytic anemia (antibody-mediated destruction of RBCs) is
    suspected. A positive Coombs test indicates that an immune
    mechanism is attacking the patient's own RBC's.
     This mechanism could be autoimmunity, alloimmunity or a drug-
    induced immune-mediated mechanism. BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    Examples of alloimmune hemolysis

    • Hemolytic disease of the newborn (also known as


    HDN or erythroblastosis fetalis)
    – Rh D hemolytic disease of the newborn (also known as
    Rh disease)
    – ABO hemolytic disease of the newborn (the indirect
    Coombs test may only be weakly positive)
    – Anti-Kell hemolytic disease of the newborn
    – Rh c hemolytic disease of the newborn
    – Rh E hemolytic disease of the newborn
    – Other blood group incompatibility (RhC, Rhe, Kidd, Duffy,
    MN, P and others)

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Examples of autoimmune hemolysis

    Warm antibody autoimmune hemolytic anemia


    ◦ Idiopathic(a disease/condition which arises spontaneously or for
    which cause unknown)
    ◦ Systemic lupus erythematosus
    ◦ Evans' syndrome (antiplatelet antibodies and hemolytic
    antibodies)

    Cold antibody autoimmune hemolytic anemia


    ◦ Idiopathic cold hemagglutinin syndrome
    ◦ Infectious mononucleosis

    Paroxysmal cold hemoglobinuria (rare)


    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    Drug-induced immune-mediated hemolysis

    • Methyldopa (IgG mediated type II


    hypersensitivity)
    • Penicillin (high dose)
    • Quinidine (IgM mediated activation of classical
    complement pathway and Membrane attack
    complex, MAC)

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Laboratory method
    The patient's red blood cells
    (RBCs) are washed (removing
    the patient's own serum) and
    then incubated with antihuman
    globulin (also known as Coombs
    reagent).
     If immunoglobulin or
    complement factors have been
    fixed on to the RBC surface in-
    vivo, the antihuman globulin will
    agglutinate the RBCs and the
    direct Coombs test will be
    positive. BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    Indirect Coombs test (ICT)
    Indirect antiglobulin test
    ~ used to detect in-vitro antibody-antigen reactions.
    ~ It is used to detect very low concentrations of antibodies
    present in a patient's plasma/serum prior to a blood
    transfusion.

     Antenatal antibody screening


    ~ used to screen pregnant women for IgG antibodies that
    are likely to pass through the placenta into the fetal blood
    and cause haemolytic disease of the newborn.
    ~ The IAT can also be used for compatibility testing, antibody
    identification, RBC phenotyping, and titration studies.
    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    Clinical uses of the indirect Coombs test

    Blood transfusion preparation


    Antibody screening
    ~ A blood sample from the recipient and a blood sample from
    every unit of donor blood are screened for antibodies with the
    indirect Coombs test.

    Cross matching
    ~ The indirect Coombs test is used to test a sample of the recipient's
    serum against a sample of the blood donor's RBCs. This is sometimes
    called cross-matching blood.

    Antenatal antibody screening


    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    First stage
    Laboratory method
    Washed test red blood cells (RBCs) are
    incubated with a test serum. If the serum
    contains antibodies to antigens on the RBC
    surface, the antibodies will bind onto the
    surface of the RBCs.

    Second stage

    The RBCs are washed three or four times with


    isotonic saline and then incubated with
    antihuman globulin. If antibodies have bound
    to RBC surface antigens in the first stage, RBCs
    will agglutinate when incubated with the
    antihuman globulin (also known Coombs
    reagent) in this stage, and the indirect Coombs
    test will be positive.
    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    Clinical uses
    • Common clinical uses of the Coombs test
    include the preparation of blood for
    transfusion in cross-matching, screening for
    atypical antibodies in the blood plasma of
    pregnant women as part of antenatal care,
    and detection of antibodies for the diagnosis
    of immune-mediated haemolytic anemias.

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE


    Coombs reagent
    Coombs antiglobulin or antihuman globulin)

    ~is used in both the direct Coombs test and the indirect
    Coombs test.
    ~It is made by injecting human globulin into animals,
    which produce polyclonal antibodies specific for human
    immunoglobulins and human complement system
    factors.
    ~More specific Coombs reagents or monoclonal
    antibodies can be used
    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
    REFERENCES
    • Denise M. Harmening PhD MLS(ASCP)
    CLS(NCA),2012.Modern Blood Banking &
    Transfusion Practices 6th Edition.by F.A.Davis
    company united sate of America.
    • Eva D.Quinley Ms MT
    (ASCP)SBB.2010.immunohaematology Principles and
    practice,3rd ed( Lippincott Williams and Wikins)
    • Jeffrey Mc Cullough.2011.Trnasfusion Medicine 3 rd
    ed,wiley Blackwell publication ,United states.
    • Harvey G.Klein,David J Anstee 2013,Mollisons blood
    transfusion in clinical medicine,12 th ed,Wiley-Blackwell.
    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE 17
    Thank you

    BIOMEDICAL SCIENCE, FACULTY OF MEDICINE 18

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