Genotyping of Different Pestiviruse Isolates by RT-PCR

Abstract

1320 blood samples were collected from herds showing clinical signs of pestiviral infections. 39 samples were positive in Ag-ELISA assay. All these 39 samples in addition to 5 cytopathic strains were cultured in MD-BK cell line and presence of pestiviral antigens was confirmed by direct-immunofluorescent test. 27 out of 44 BVDV suspected isolates were detected by RT-PCR using a primer set. Differentiation among the viruses was achieved by cutting the PCR products with restriction endonucleses enzymes Ava1, Bgl1 and Alu1. Using this procedure it was possible to distinguish at least two genogroups, 1 and 2 BDV containing 14 and 3 isolates, respectively.