An enzyme-linked immunosorbent assay (ELISA) for antibodies to bovine viral diarrhea virus.
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.
Durham
P J
PJ
Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
Hassard
L E
LE
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000096:biosynthesis / Q000276:immunology; D002417:Cattle; D002460:Cell Line; D001908:Diarrhea Viruses, Bovine Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007231:Infant, Newborn; D009500:Neutralization Tests; D010576:Pestivirus / Q000276:immunology*; D011237:Predictive Value of Tests; D015203:Reproducibility of Results
D000914:Antibodies, Viral; D000956:Antigens, Viral