Detection of bovine gammaherpesviruses by a nested duplex PCR

The two bovine gammaherpesviruses (Alcelaphine herpesvirus 1, AlHV-1; Bovine herpesvirus 4, BoHV-4) are distributed worldwide in cattle populations. Since the animals are frequently infected latently with no or low seropositivity, a DNA based diagnostic method would be useful for surveys and detection of these viruses. In the present study a nested duplex PCR was established for the sensitive and specific simultaneous detection of both viruses. The primers were designed for the gene of the major capsid protein (ORF25). The assay did not amplify the capsid gene sequence of 10 related bovine herpesviruses and other gammaherpesviruses. The test was able to detect 1 pfu (plaque forming unit) of AlHV-1 and BoHV-4. Among 146 clinical samples (lymph nodes and peripheral blood leukocytes, PBLs) 65 (44.52%) were found to be positives for AlHV-1 and 84 (57.53%) for BoHV-4. This nested duplex PCR assay could serve as a useful diagnostic method for rapid, sensitive, specific simultaneous detection of the two bovine gammaherpesviruses.