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    in the presence of electrolytes at a suitable temperature and Ph,

     Qualitative agglutination test


     Agglutination tests can be used in a qualitative manner to assay for the presence
    of an antigen or an antibody. The antibody is mixed with the particulate antigen
    and a positive test is indicated by the agglutination of the particulate antigen.
     For example, to determine patient’s blood type the red blood cells of the person
    can be mixed with antibody to a blood group antigen. Another example is that to
    assay the presence of antibodies in a sample, the serum of the patient is mixed
    with the red blood cell (RBC) of a known blood type.

     Quantitative agglutination test


     To measure the level of antibodies to particulate antigens, agglutination test can
    be widely used. For this test, serial dilutions of the sample can be made and it is
    tested for antibody. Then a fixed amount of particulate antigen or bacteria or red
    blood cells can be added to it. Determine the maximum dilution which forms
    agglutination and this maximum dilution which gives observable agglutination is
    known as the titer. The results is shown as the reciprocal of the maximum dilution
    that forms visible agglutination.
    SLIDE AGGLUTINATION TEST

     Used for serotyping (e.g. Salmonella)


     Antigen: isolated Salmonella in suspension
     Antibody: specific antisera against Salmonella
     Place test Salmonella in a drop of saline on a slide
     Add a drop of antiserum, mix and rock slide for approx. 1 minute
     Examine for agglutination

    6
    SLIDE AGGLUTINATION TEST
    7
    TUBE AGGLUTINATION
    TEST
     Also known as the standard agglutination test or serum agglutination test (SAT)
     Test serum is diluted in a series of tubes (doubling dilutions)
     Constant defined amount of antigen is then added to each tube and tubes
    incubated…
     Particular antigen clumps at the bottom of the test tube
     Test is read at 50% agglutination
     Quantitative
     Confirmatory test for ELISA reactors
     Example: Brucellosis screening , Widal Testing

    8
    TUBE AGGLUTINATION TEST
    Agglutination No agglutination

    1/10 1/20 1/40 1/80 1/160 1/320 Neg. ctrl

    In this case, the titre is 1/40


    Soluble antigen

    particulated antigen
    PASSIVE AGGLUTINATION

     Hemagglutination, or haemagglutination, is a specific form of agglutination


    that involves red blood cells (RBCs). It has two common uses in the laboratory:
    blood typing and the quantification of virus dilutions in a heamagglutination
    assay.
     EX: detection of illegal drugs.
     Latex agglutination is observed when a sample containing the specific
    antigen (or antibody) is mixed with an antibody (or antigen) which is coated on
    the surface of latex particles.
     EX: detection of pregnancy, ASOT.(antigen of streptococcus O- antigen).
    PROZONE EFFECT
     Occasionally, it is observed that when the concentration of
    antibody is high, there is no agglutination and then, as the
    sample is diluted, agglutination occurs.
     The lack of agglutination at high concentrations of antibodies
    is called the prozone effect.
     Lack of agglutination in the prozone is due to antibody excess
    resulting in very small complexes that do not clump to form
    visible agglutination.

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