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Updated: May 30, 2026
Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids
Published on: March 17, 2016
Development of a sandwich ELISA for the detection of bovine herpesvirus type 1
Shanaz Bashir1, Rashmi Singh, Barkha Sharma
1Department of Epidemiology and Vety. Preventive medicine, College of Veterinary Sciences, Veterinary University, Mathura (UP), India.
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 1:4 000, whereas the conjugate was used at 1:2 000 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.
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