Rapid and sensitive PCR-based test for detection of Bovine herpesvirus-1 in semen

A polymerase chain reaction (PCR) based rapid and sensitive detection test was established for detection of bovine herpesvirus-1 (BHV-1) using primers designed from glycoprotein C gene. PCR reaction was optimized with DNA extracted from BHV-1-infected cell culture supernatant to amplify a specific 520 bp amplicon from BHV-1 DNA. The amplified 520 bp PCR product was characterized using restriction endonucleases and Southern blot hybridization. The sensitivity of PCR was found to detect as low as 10⁻¹ plaque forming unit (pfu) BHV-1. The test developed was applied to frozen bovine semen artificially spiked with BHV-1. The sensitivity of detection was as low as 10⁻¹ plaque forming unit (pfu) BHV-1.