A POLYMERASE CHAIN-REACTION (PCR) ASSAY FOR THE DETECTION OF BOVINE HERPESVIRUS-1 (BHV1) IN SELECTIVELY DIGESTED WHOLE BOVINE SEMEN

A PCR assay for the detection of bovine herpesvirus type 1 (BHV1) DNA in selectively digested whole bovine semen was developed and evaluated . A brief treatment with proteinase-K was used to lyse free virus, vir us present in non-sperm cells and virus adhering to the spermatozoa. G enomic bovine DNA was not released by this treatment. Primers and prob es were based on the nucleotide sequence of the go gene. BHV1 virus-sp iked split samples were used as positive controls and the PCR products were detected by eye in ethidium bromide-stained agarose gels. Sequen tially collected non-extended semen samples from experimentally infect ed bulls were used to compare this assay with virus isolation. Of a to tal of 162 ejaculates, 51 were found positive by virus isolation, wher eas PCR detected BHV1 DNA in 73. PCR detected BHV1 DNA for a longer pe riod after infection and reactivation. Apart from its superior sensiti vity, this PCR assay also has the advantage of being a relatively simp le procedure, providing results within 24 h.