COMPARISAN OF THE TECHNIQUESS OF VIRAL ISOLATION AND NESTED PCR IN THE DETECTION of BHV-1 IN BOVINE SEMEN CONTAMINATED EXPERIMENTALLY AND NATURALLY

ABSTRACTS

Bovine Herpesvirus type 1 (BHV-1) is the causal agent of Bovine Infectious Rinotracheitis (IBR), (IPV) and (IBP) and it is the main viral pathogen found in bovine semen. The gold standard for BHV-1 direct diagnosis in bovine semen samples is viral isolation, which has numerous disadvantages. The aim of this study was to evaluate a nested PCR (nPCR) assay for BHV1 detection in bovine semen. The primers targeting gB and the specify of the amplicons was confirmed with Fnu4HI and Sau96I restriction profile. The nPCR showed fast and with a high analytic sensibility (10⁵ times higher than viral isolation). When the assay was applied to a collection of semen samples collected from a bull from an Artificial Insemination Center with a single semen sample positive at viral isolation, the nPCR was able to detect the BHV-1 DNA 27 days before and 3 days after the viral isolation.

KEY WORDS:
IBR; semen; PCR.