Monoclonal antibody capture enzyme-linked immunosorbent assay for detection of bovine enteric coronavirus

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Abstract
Monoclonal antibodies reactive with 3 different viral polypeptides were evaluated singly and in combination as the capture antibody(s) in an ELISA system for the detection of bovine enteric coronavirus. Similar levels of sensitivity were found for all combinations tested. A sensitive, highly specific and reproducible assay for the detection of bovine enteric coronavirus was developed, using a mixture of 2 of these monoclonal antibodies reactive with antigenic components external or internal to the virion. These monoclonal antibodies were bound indirectly to 96-well plates via rabbit anti-mouse Ig. After sample application and incubation, virus was detected by using rabbit anti-coronavirus peroxidase conjugate followed by enzyme substrate and chromagen. Fecal samples from a single herd of cows were screened for the presence of coronavirus by this assay. Of clinically normal cows, 5% were shedding coronavirus.