Development of a sandwich ELISA for the detection of bovine herpesvirus type 1

ABSTRACT

OBJECTIVE@#To develop a standard enzyme-linked immunosorbent assay (ELISA) for the detection of bovine herpesvirus type 1 (BHV-1).@*METHODS@#The assay was based on hyperimmune rabbit and guinea pig antisera raised against purified BHV-1. Polyethylene glycol precipitation and sucrose density gradient methods were adopted for viral concentration and purification. Antisera were raised using Freund's adjuvant followed by extraction of IgG of high purity.@*RESULTS@#Optimum antisera dilutions as determined by titrations were chosen as 14 000, whereas the conjugate was used at 12 000 dilution. Using 95 clinical specimens, the ELISA test showed a sensitivity and specificity of 91.90 % and 93.10 %, respectively when compared to PCR. The cut-off value was fixed at 0.15 (A(490)) and a P/N ratio of >1.30 indicated a significant positive reaction.@*CONCLUSIONS@#The results have demonstrated that this ELISA could efficiently detect BHV-1 and can be used as an important diagnostic tool.