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Immunological testing

Immunological testing. Monoclonal antibodies Serology Quantifying antigen – antibody reactions. Perspective 17.1 Monoclonal Antibodies. Serology. Antibodies Antibodies detect and identify antigens. Quantifying antigen – antibody reactions. Seroconversion or rise in titer

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Immunological testing

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  1. Immunological testing • Monoclonal antibodies • Serology • Quantifying antigen – antibody reactions

  2. Perspective 17.1 Monoclonal Antibodies

  3. Serology • Antibodies • Antibodies detect and identify antigens

  4. Quantifying antigen – antibody reactions • Seroconversion or rise in titer • Serial dilutions

  5. Figure 17.2 - Quantitation of immunologic tests

  6. Precipitation reactions • Immunodiffusion • Immunoelectrophoresis

  7. Figure 17.3 Precipitation reaction

  8. Figure 17.4 Immunodiffusion

  9. Figure 17.6 Immunoelectrophoresis

  10. Agglutination reactions • Direct agglutination • Indirect agglutination • Hemagglutination

  11. Direct agglutination • Cross – linking and lattice formation • Antibodies react with particulate antigens (red blood cells, bacteria, fungi) • Visible clumps • Estimate amount of antibody

  12. Indirect agglutination • Soluble antigen is coated onto particles (red bood cells, latex beads) • Allow for visible clumps (agglutination)

  13. Figure 17.7 - Agglutination reaction

  14. Immunofluorescence tests • Direct fluorescent antibody test • Indirect fluorescent antibody test

  15. Figure 17.8 - Direct and indirect fluorescent antibody test

  16. Antigen – antibody assays • Radioimmunoassay (RIA) • Enzyme – linked immunosorbant assay (ELISA) • Western blot

  17. Radioimmunoassay (RIA) • Competitive inhibition assay • Measure antigen or antibody • Ex. Measure small amounts of hormones or drugs in a clinical sample • Ex. Measure small amounts of IgE antibody (radioallergosorbent test)

  18. Enzyme – linked immunosorbant assay (ELISA) • Color reaction assay • Indirect ELISA • Direct ELISA

  19. Figure 17.9 - Indirect ELISA

  20. Figure 17.1 - Direct ELISA

  21. Figure 17.1 Western blot

  22. Complement fixation test • Measures the binding of complement by an antigen – antibody interaction • Indicator system determine positive or negative reactions

  23. Figure 17.1Complement fixation test

  24. Neutralization test • Antibody bind to specific antigen (virus, toxin) • Antibody – antigen complex prevents antigen from binding (neutralization)

  25. Cellular immunology test • Identification of subsets of lymphocytes • Lymphocyte response to mitogens • Cytoxic T – cell function • Cell – mediated immunity to infectious agents

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