Detection and genotyping of bovine diarrhea virus by reverse transcription-polymerase chain amplification of the 5 ' untranslated region

A reverse-transcription polymerase chain reaction (RT-PCR) was developed to differentiate the bovine diarrhea virus (BVDV) from other pestiviruses, an d to determine the genotype of the BVDV isolates. For this purpose, primer pairs were selected in the 5' untranslated region (5'UTR). The primers B-E and B-2 were located in highly conserved regions and were pestivirus-specif ic. Two primer pairs named B3B4 and B5B6 were specific of BVDV genotypes I and II, respectively. With this technique, an amplification product of the expected size was obtained with either the B3B4 or the B5B6 primer pairs fo r the 107 BVDV isolates tested but not for BDV or CSFV. For some isolates t hat were grouped in the genotype II, sequence analysis of the PCR fragments confirmed their classification into this genotype. (C) 1999 Elsevier Scien ce B.V. All rights reserved.