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Immunologic Laboratory Tests

Immunologic Laboratory Tests. Kristine Krafts, M.D. Immunologic Lab Tests Outline. Agglutination reactions DAT IAT Immunofluorescence ELISA Western blot Flow cytometry. Things to Remember About Each Test. What does it measure? Where does the Ag-Ab interaction occur?

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Immunologic Laboratory Tests

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  1. Immunologic Laboratory Tests Kristine Krafts, M.D.

  2. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence • ELISA • Western blot • Flow cytometry

  3. Things to Remember About Each Test • What does it measure? • Where does the Ag-Ab interaction occur? • How is the Ag-Ab complex detected?

  4. Immunologic Lab Tests Outline • Agglutination reactions

  5. Agglutination Reactions: Purpose • Detection of Ag in the patient’s specimen • Examples: • testing for antibodies to infectious agents • testing for Hemophilus influenzae type B capsular antigen in CSF

  6. negative positive

  7. Immunologic Lab Tests Outline • Agglutination reactions • DAT

  8. DAT: Purpose • Detection of Ab on patient’s red cells • Also called the direct Coombs Test • Performed in patients with hemolytic anemia

  9. patient red cells + AHG = agglutination

  10. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT

  11. IAT: Purpose • Detection of antibodies to red cell antigens • Also called the indirect Coombs Test • Performed as part of pre-transfusion testing

  12. patient Ab donor RBC Ab-coated donor RBC + = + AHG agglutination =

  13. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence

  14. Immunofluorescence: Purpose • Detection of a specific antigen or antibody in a patient’s specimen • Examples: • detection of antigen in a patient’s biopsy specimen • detection of antibody in a patient’s serum

  15. Positive fluorescent antinuclear antibody test

  16. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence • ELISA

  17. ELISA: Purpose • Detection of antibodies in patient specimen • Examples: • home pregnancy tests • HIV tests • tests for some coagulation factors, cytokines, and autoantibodies

  18. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence • ELISA • Western blot

  19. Western Blot: Purpose • Detection of antibodies in patient specimen • Most common example: HIV test

  20. Enough bands = patient is “positive”

  21. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence • ELISA • Western blot • Flow cytometry

  22. Flow Cytometry: Purpose • Characterization of cell size, complexity, antigens • Examples: • diagnosis of leukemia and lymphoma • determination of CD4/CD8 counts in patients with HIV

  23. Immunologic Lab Tests Outline • Agglutination reactions • DAT • IAT • Immunofluorescence • ELISA • Western blot • Flow cytometry

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