Enzyme-linked immunosorbent assays based on polyclonal and monoclonal antibodies for rotavirus detection

Abstract

We describe two enzyme-linked immunosorbent assays for rotavirus antigen in feces, which were designed to be as sensitive and specific as possible, and easy to use anywhere. Both are indirect methods, using the antibody capture method, but the second assay utilizes a rotavirus group-specific monoclonal 'detecting' antibody instead of the hyperimmune polyvalent guinea pig antisera used in the first assay. Both tests were found to be more sensitive than electron microscopy for detecting virus. To develop these tests, solid phase, antiserum production methods, treatment of the test antigen with EDTA, substrate, stability of reagents, and the need for confirmatory 'blocking' tests were all examined. The first assay described is that used at present by the World Health Organization for their worldwide diarrheal disease control program.