An enzyme-linked immunosorbent assay (ELISA) with high sensitivity was developed for use in the avian system. Anti-reovirus antibody produced in chickens against the isolants S1133, UMI-203, Reo-25, and Winterfield were detected with this method at high titers (up to 1:14,000).
A microplate enzyme-linked immunosorbent assay was developed to detect chicken anti-reovirus 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 1:51,000, 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. According to Schuurs and van Weemen (13), the number of publications concerning various
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