Observe figure 17.15 and make note of the several steps in the indirect ELISA test. What four
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(a) Indirect ELISA, comparing a positive vs. negative reaction. This is the basis for HIV screening tests. (b) Microtiter ELISA Plate with 96 Tests for HIV Antibodies Colored wells indicate a positive reaction. Well Well Known antigen is adsorbed to well Serum samples with unknown A antibodies Well is rinsed to remove unbound (c) Capture or Antibody Sandwich ELISA Method. Note that an antigen is trapped between two antibodies. This test is used to detect the measles virus. Antibody specific to test antigen is adsorbed to well. Indicator antibody attaches to any bound antibody Test antigen is added antigen binds to antibody Wells are rinsed to remove unbound indicator antibody A colorless substrate for enzyme is added. Enzyme-linked antibody specific for test antigen then binds to different antigenic site forming a sandwich. Enzymes linked to indicator Ab hydrolyze the substrate, which releases a dye Wells that develop color are positive for the antibody colorless wells are negative. Enzyme's substrate (2) is added, and reaction produces a visible color change ()
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Related Book For
Foundations in Microbiology
ISBN: 978-0073375298
8th edition
Authors: Kathleen Park Talaro, Barry Chess
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