Isolation of BHV-1 from bovine semen and application of real time PCR for diagnosis of IBR/IPV from clinical samples.

Abstract

Five hundred and twenty three semen samples were collected from fi fteen different semen collection centres located across four states of South India. These samples were screened for infectious bovine rhinotracheitis virus by virus isolation studies at the Institute of Animal Health and Veterinary Biologicals, Bangalore, India. Bovine turbinate (BT) and Madin Darby Bovine Kidney (MDBK) cell lines were used for virus isolation. Out of the total samples, four semen samples showed cytopathic changes in cells and were further confi rmed by regular PCR targeting amplifi cation of the gC region of bovine herpesvirus-1. Real time PCR was applied for these virus isolates using primers and probes amplifying the gB region of bovine herpesvirus-1. Further, this real time PCR protocol was adapted for diagnosis of IBR/IPV directly on the clinical samples.