A Qualitative PCR Assay for the Discrimination of Bubaline Herpesvirus 1, Bovine Herpesvirus 1 and Bovine Herpesvirus 5

Bubaline herpesvirus 1 (BuHV-1), Bovine herpesvirus 1 (BoHV-1) and Bovine herpesvirus5 (BoHV-5) are classified in the genusVaricellovirus, subfamilyAlphaherpesvirinae. BoHV-1 is thecausative agent of infectious bovine rhinotracheitis, BoHV-5 induces moderate disease in adult cattlewhile BuHV-1 has instead been associated with a decline in livestock production of water buffaloes.The aim of this study was to develop a qualitative PCR assay that allows the discrimination ofBuHV-1, BoHV-1 and BoHV-5. The alignment of homologous genes identified specific nucleotidesequences of BuHV- 1, BoHV-1 and BoHV-5. The design of the primers and the optimization of thePCR assay were focused on the target sequences located on the portions of gD, gE and gG genes.This assay involved the use of three different PCR end-points: the PCR of a portion of the gD geneidentified only the presence of BoHV-1; the PCR of a portion of the gE gene confirmed the presenceof both BoHV-5 and BuHV-1; the PCR of a portion of the gG gene discriminated between BoHV-5and BuHV-1, as the amplification product was observed only for BoHV-5. This qualitative PCR assayallowed the differentiation of BoHV-1 and BoHV-5 infections both in cattle and water buffaloes andheterologous BuHV-1 infections in bovine.