THE DISTINCTION OF SEROLOGICALLY RELATED RUMINANT ALPHAHERPESVIRUSES BY THE POLYMERASE CHAIN-REACTION (PCR) AND RESTRICTION-ENDONUCLEASE ANALYSIS

The amplification and analysis of a 468bp fragment from the gB gene of the serologically related ruminant alphaherpesviruses bovine herpesvi rus-1.1 and 1.2 (BHV-1.1 and BHV-1.2), caprine herpesvirus-1 (CapHV-1) , cervine herpesvirus-1 (CerHV-1) and rangiferine herpesvirus-1 (RanHV -1) by PCR and restriction endonuclease analysis is described. As prim ers, 22bp oligomers selected from the BHV-1 gB gene sequences were use d for the amplification of the DNA from the five viruses. The amplific ation product from each virus was analysed by the restriction endonucl ease enzymes BglI, HinfI, SmaI and AvaI. The specific amplification ob tained demonstrate the existence of the gB gene sequences for each of the five alphaherpesviruses. However, sequences from some of the fragm ents were found to be different from those predicted from the gB gene following restriction endonuclease analysis. All five amplification pr oducts generated the same number of fragments after digestion with Hin fI except for two additional bands evident in CapHV-1. The CerHV-1 and RanHV1 fragments contained slightly different BglI restriction sites from those of the other three. While BHV-1.1, BHV-1.2, CapHV-1 and Cer HV-1 contained SmaI and AvaI restriction sites, the RanHV-1 amplificat ion product lacked both SmaI and AvaI restriction sites.