Adaptation of enzyme-linked immunosorbent assay to the avian system.
A microplate enzyme-linked immunosorbent assay was developed to detect chicken anti-rovirus antibodies. Studies of the parameters which affect the outcome of the assay with avian serum revealed two aspects for a successful assay. First, enzyme-antibody conjugates prepared by the periodate oxidation technique were found to have retained far more immunological activity than conjugates produced by a glutaraldehyde cross-linking. Second, the results indicated an unusually high affinity of chicken immunoglobulin for the microplate plastic which was mostly eliminated by a pretreatment technique with fixed fetal calf serum. The enzyme-linked immunosorbent assay compared favorably with the latex passive agglutination test, yielding a titration endpoint of 1:511,000, or approximately 1,300 times more sensitive than the latex passive agglutination assay. The assay proved not only to be sensitive to less than 1 ng of specific antibody, but also to have low to moderate variance and high reliability.
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D001717:Birds / Q000276:immunology* / Q000382:microbiology; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D007136:Immunoglobulins / Q000032:analysis; D007841:Latex Fixation Tests; D012087:Reoviridae / Q000276:immunology
D000914:Antibodies, Viral; D007136:Immunoglobulins