免疫学检测与免疫学技术.pptVIP

Ag-Ab reactionsTests for Ag-Ab reactions Nature of Ag/Ab Reactions Calculation of Affinity Specificity The ability of an individual antibody combining site to react with only one antigenic determinant. The ability of a population of antibody molecules to react with only one antigen. Tests Based on Ag/Ab Reactions All tests based on Ag/Ab reactions will have to depend on lattice formation or they will have to utilize ways to detect small immune complexes All tests based on Ag/Ab reactions can be used to detect either Ag or Ab Agglutination Tests Agglutination/Hemagglutination Quantitative agglutination test Titer Prozone Agglutination/Hemagglutination Definition Qualitative test Quantitative test Passive Agglutination/Hemagglutination Definition - agglutination test done with a soluble antigen coated onto a particle Coombs (Antiglobulin)Tests Coombs (Antiglobulin)Tests Indirect Coombs Test Detects anti-erythrocyte antibodies in serum Coombs (Antiglobulin)Tests Applications Detection of anti-Rh Ab Autoimmune hemolytic anemia Agglutination/Hemagglutination Inhibition Definition - test based on the inhibition of agglutination due to competition with a soluble Ag Agglutination/Hemagglutination Inhibition Applications Measurement of soluble Ag Practical considerations Same as agglutination test Precipitation Tests Lattice Formation Radial Immunodiffusion (Mancini) Interpretation Diameter of ring is proportional to the concentration Quantitative Ig levels Immunoelectrophoresis Method Ags are separated by electrophoresis Immunoelectrophoresis Method Interpretation Qualitative Relative concentration Countercurrent electrophoresis Method Ag and Ab migrate toward each other by electrophoresis Used only when Ag and Ab have opposite charges Radioimmuoassays (RIA)Enzyme-Linked Immunosorbent Assays (ELISA) Lattice formation not required Competitive RIA/ELISA for Ag Method Determine amount of Ab needed to bind to a known amount of labeled Ag Competitive RIA/ELISA for Ag