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Serology. Eva L. Dizon, M.D. Microbiology Our Lady of Fatima University. Serology. Antigen + Antibody. Immune Complex. Test that depends on the Formation of Immune Complex Nephelometry Immunodiffusion Complement fixation Cryoglobulins.

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    1. Serology Eva L. Dizon, M.D. Microbiology Our Lady of Fatima University

    2. Serology Antigen + Antibody Immune Complex

    3. Test that depends on the Formation of Immune Complex Nephelometry Immunodiffusion Complement fixation Cryoglobulins Test in which Ag + Ab is fixed to a solid surface Agglutination Elisa Electrophoresis Serology

    4. Amount of Precipitine Antigen Excess (Postzone) Antibody Excess (Prozone) Equivalence Increasing Antigen Concentration

    5. Immunodiffusion A A A Present in reduce concentration I B B B Zone of equivalence + precipitation I Contaminant present in antigen I I

    6. Ouchterlony • Double diffusion • Characterize relationship between 2 antigen • If antigen are in the same precipitation line ,they fail to cross

    7. Ouchterlony Ag 1 Ag 2 Ag 4 Ab Limitations : Insensitive Needs large unit of Ab and Ag Resolve by CIE –placing agar in electric field Ag 3

    8. Nephelometry • IC in solution is monitored by spectrometry • Formation of IC is measured in a photoelectric cell as optical density Add increasing mount of antigen Ag Ag Ag

    9. Complement fixation test IC is fixed and consume complement 1. IC (ag +ab) + complement 2. IC +C +RBC------ no hemolysis (positive) Ag + ____ + C +RBC -------- hemolysis (negative)

    10. Cryoglobulins This are serum globulin that precipitate at T less than 37C To detect the presence in the blood ---- serum Samples are incubated in 4C for several hours ----- note for precipitation reaction Precipitated proteins isolated by centrifugation then assayed for IG by nephelometry,electrophoresis

    11. Ag and Ab fixed to a solid surface Agglutination test for the presence of antibody Indirect (Passive) Particle RBC Ag Ag Ag + Ab Ag Particle agglutination

    12. Neutralization test Hemmagglutination inhibition test Virus + RBC -------- Agglutinate Virus+ Ab + RBC -------- No Agglutination

    13. ASO titer Streptolysin O (Ag) + Serum of patient Agglutination If Ptx had Strep pyogenes infection Ab against Streptolysin O is present

    14. ELISA Direct – look for antigen Slide with Known ab ------ put serum ?Ag -------wash--------add enzyme labeled antibody--------wash--------look under spectrophotometer

    15. Elisa Indirect – look for antibody Slide with antigen --------+? Ab --------wash---------+ antihuman globulin -----------wash-------- look under spectrophotometer

    16. Fluoresence test Direct- looks for antigen 1. Slide put an antigen 2. Add fluorescein labeled antibody 3. Examine under the UV microscope Indirect- antibody Slide with known antigen Wash Add fluorescein labeled antihuman Ab

    17. Thank you and read your Manual TOO!