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    Serological Methods For Lab Diagnosis 2

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    Devender Rana
    Serology is the scientific study of blood serum to detect antibodies and antigens through in vitro antigen-antibody reactions. Various serological methods can be used including precipitation, agglutination, immunoelectrophoresis, complement fixation, enzyme-linked immunosorbent assay, and immunofluorescence assay. Enzyme-linked immunosorbent assay (ELISA) is commonly used and can detect antibodies or antigens to indicate any infectious disease, provided the appropriate antibodies and antigens are available. Sensitivity and specificity are important factors in evaluating serological test methodology.

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    Serological Methods For Lab Diagnosis 2

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    Devender Rana
    59 views4 pages
    Serology is the scientific study of blood serum to detect antibodies and antigens through in vitro antigen-antibody reactions. Various serological methods can be used including precipitation, agglutination, immunoelectrophoresis, complement fixation, enzyme-linked immunosorbent assay, and immunofluorescence assay. Enzyme-linked immunosorbent assay (ELISA) is commonly used and can detect antibodies or antigens to indicate any infectious disease, provided the appropriate antibodies and antigens are available. Sensitivity and specificity are important factors in evaluating serological test methodology.

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    Serological Methods for Lab Diagnosis 2

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    Serology is the scientific study of blood serum to detect antibodies and antigens through in vitro antigen-antibody reactions. Various serological methods can be used including precipitation, agglutination, immunoelectrophoresis, complement fixation, enzyme-linked immunosorbent assay, and immunofluorescence assay. Enzyme-linked immunosorbent assay (ELISA) is commonly used and can detect antibodies or antigens to indicate any infectious disease, provided the appropriate antibodies and antigens are available. Sensitivity and specificity are important factors in evaluating serological test methodology.

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    59 views4 pages

    Serological Methods For Lab Diagnosis 2

    Uploaded by

    Devender Rana
    Serology is the scientific study of blood serum to detect antibodies and antigens through in vitro antigen-antibody reactions. Various serological methods can be used including precipitation, agglutination, immunoelectrophoresis, complement fixation, enzyme-linked immunosorbent assay, and immunofluorescence assay. Enzyme-linked immunosorbent assay (ELISA) is commonly used and can detect antibodies or antigens to indicate any infectious disease, provided the appropriate antibodies and antigens are available. Sensitivity and specificity are important factors in evaluating serological test methodology.

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    of 4

    SEROLOGICAL

    METHODS FOR LAB


    DIAGNOSIS

    BY
    DEVENDER SINGH RANA
    2006V19B
    Serology-The branch of laboratory medicine that studies blood
    serum for evidence of infection and other parameters by
    evaluating antigen-antibody reactions in vitro. OR
    Serology is the scientific study of blood serum. In practice, the
    term usually refers to the diagnostic identification of antibodies
    in the serum .
    We can detect antigens too.

    Various serological methods may be-


    1. Precipitation
    2. Agglutination
    3. Imunoelectrphoresis (IEP)
    4. Complement fixation (CF)
    5. Enzyme-linked immunosorbent assay
    6. Immunofluorescence Assay

    1.Precipitation
    Principle- In this soluble antigen and antibody (in proper
    proportions) form visible precipitate .Ther is lattice formation if
    antigen binds with Fab sites of 2 antibodies.
    Various precipitation methods are:
    – Double diffusion (Ouchterlony)
    – Single diffusion (radial immunodiffusion)
    – Imunoelectrphoresis
    – Immunofixation

    2.Agglutination
    Principle- In this particulate antigen and antibody combine to form
    clumping .There is lattice formation if antigen binds with Fab sites
    of 2 antibodies forming bridges between antigens.
    Various agglutination methods may be-

    – Direct agglutination (Blood Bank)


    – Passive Hemagglutination (treat RBC's with tannic acid
    to allow adsorption of protein antigens)
    – Passive latex agglutination (antigen attached to latex
    particle)

    3.Imunoelectrphoresis (IEP)
    Qualitative- In this a serum sample is electrophoresed through an
    agar medium.
    A trough is cut in the agar and filled with Ab.A precipitin arc is
    then formed because Ag diffuses radially and Ab from a trough
    diffuses, the reactants meet in optimal proportions for
    precipitation.

    4.Complement fixation (CF) Test – In this


    antibody and antigen allowed to combine in presence of
    complement .
    If complement is fixed by specific antigen-antibody reaction, it
    will be unable to combine with indicator system
    Precautions
    1. Serum must be heat-activated
    2. Stored serum becomes anti-complementary
    3. Extensive QC/standardization required
    4. Only use for IgM antibodies

    5.Enzyme-linked immunosorbent assay- It is also called


    Enzyme-linked immunosorbent assay or ELISA, enzyme
    immunoassay or EIA, is a biochemical technique used mainly in
    immunology to detect the presence of an antibody or an antigen in
    a sample. The ELISA has been used as a diagnostic tool in
    medicine .Because the ELISA can be performed to evaluate either
    the presence of antigen or the presence of antibody in a sample .
    ELISA methods can detect any infectious disease provided if we
    have antibodies and antigen to any infection, enzyme or any
    substance.

    Some common terms used in evaluating test methodology


    are-
    1.Sensitivity
    Analytical Sensitivity – ability of a test to detect very small
    amounts of a substance .
    Clinical Sensitivity – ability of test to give positive result if
    patient has the disease (no false negative results) .

    2. Specificity
    Analytical Specificity – ability of test to detect substance without
    interference from cross-reacting substances .
    Clinical Specificity – ability of test to give negative result if patient
    does not have disease (no false positive results) .

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