| Summary Bovine  herpesvirus 1 (BoHV-1) infection in cattle and buffalo makes these animals  life-long carriers of the virus which is intermittently excreted in semen. In  the present study, a real-time polymerase chain reaction (PCR) was validated to  screen frozen semen from cattle and buffalo for BoHV-1 by amplification of the gB gene of the virus. Analysing the intra- and inter-test variability, the assay  was found to be highly reproducible. High sensitivity (100%) and specificity  (90.04%) of this real-time PCR assay was recorded in comparison to virus  isolation. Extended frozen semen samples from 574 cattle and buffalo bulls  that were seropositive to infectious bovine rhinotracheitis (IBR) tested by  real-time PCR indicated that 1.97% semen batches from cattle and 3.36% batches  of buffalo semen were positive for BoHV-1. The  real-time PCR protocol will be useful for screening large numbers of semen  samples from IBR-seropositive cattle and buffalo bulls as the test is less time  consuming and several batches of semen can be tested with ease compared to  virus isolation in cell culture.
 KeywordsBoHV-1,  Bovine herpesvirus, India, Infectious bovine rhinotracheitis, PCR, Polymerase  chain reaction, Real-time PCR, Semen, Virus.
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