GERD Exercise 12: Serological Testing > Activity 3: Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Introduction Introduction ‘The enzyme-linked immunsorbent assay (ELISA) is used to test forthe presence ofan antigen ‘orantibody. The asay is considered enzyme linked because an enzyme is chemically linked to an antibody in both the direct and indirect versions ofthe test. Immunosorbent refers tothe fact that either antigens or antibodies are being adsorbed (stuck to plastic the tests, designed to detect an antigen or antigens tis adlrect ELISA because i is directly looking for the foreign substance. An indirect ELISA i designed to detect antibodies thatthe patient has ‘made against the antigen. A postive result withthe indirect ELISA requires seroconversion. Seroconversion occurs when a patient goes fom testing negative fora specific antibody to testing positive for the same antibody. Inthe diet ELISA, 296-well microtiter plate i coated with homologous antibodies made agains the antigen of interest. The number of wells makes it easy to test many samples atthe same time. The patient serum sample's added tothe plate to test for the presence of the antigen tha binds to the antibody coating onthe plate ELISA takes advantage ofthe fac that protein sticks well to plastic. A secondary antibady is added tothe plate after the patient serum sample is added. the antigen s present, a"sandwichof antibody, antigen, and secondary antibody will form. The secondary antibody is chemically inked t an enzyme When the substrate is added, the enzyme converts the substrate from a colorless compound oa colored compound, The amount of color produced wil be proportional to the amount of antigen present binding tothe antibodies and, thus, determines whether the patients positive forthe antigen. Ifthe antigen snot present, the secondary (enzyme-linked) antibodies willbe rinsed On ©