An enzyme-linked immunosorbent assay (ELISA) for antibodies to bovine viral diarrhea virus

Abstract

A single dilution enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to bovine viral diarrhea (BVD) virus in cattle sera. Viral antigen (NADL strain) was grown in a pig kidney cell line (PK15), and after removal of nuclear debris, was purified by ultracentrifugation through a potassium tartrate cushion. Antigen grown in embryonic bovine tracheal epithelial cells was also satisfactory. The test used a high salt buffer to minimize nonspecific reactivity, polyethylene glycol to enhance the reaction, and Protein G as the labelling agent. Comparative testing with the virus neutralization test (VNT) showed the ELISA results to have a high level of correlation with the VNT titers ( r = 0.83). In vaccinated animals the ELISA detected antibodies earlier than the VNT. All animals sampled from a BVD-free herd were negative for BVD antibody. The single dilution test showed close agreement ( r = 0.84) with ELISA values obtained using a serial dilution technique, and also proved to have a high level of reproducibility. The test procedures were relatively easy to carry out, and were economic in their use of materials.