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Identification of cattle infected with bovine virus diarrhoea virus using a monoclonal antibody capture ELISA
A Fenton1, P F Nettleton, G Entrican
1Moredun Research Institute, Edinburgh, Scotland.
Abstract:
A monoclonal antibody capture enzyme linked immunosorbent assay (ELISA) has been developed to detect pestivirus-specific antigen in the leucocytes of cattle infected with bovine virus diarrhoea virus (BVDV). A blind trial was conducted to compare the specificity of the ELISA with conventional tissue culture virus isolation on 215 blood samples submitted for BVDV diagnosis from cattle throughout Scotland. One hundred and sixty seven samples were negative by both ELISA and virus isolation and 47 samples were positive by both tests. One blood was negative by ELISA and positive by virus isolation.
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01:15Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
01:33Enzyme-Linked Immunosorbent Assay
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.