REVERSE TRANSCRIPTION COMBINED WITH POLYMERASE CHAIN-REACTION AS A DETECTION METHOD FOR PESTIVIRAL INFECTIONS

An assay based on reverse transcription coupled with the polymerase ch ain reaction (RT-PCR) was used for the detection of hog cholera virus (HCV) and bovine virus diarrhoea virus (BVDV) in cell culture. In this study, a precipitate of the supernatants derived from cell cultures i nfected with HCV and BVDV was used in RT reactions, in place of extrac ted viral RNA. Both RT and PCR were performed using recombinant Thermu s thermophilus (rTth) DNA polymerase. The specificity of the RT-PCR pr oducts was confirmed by hybridisation with a digoxygenin-labelled DNA probe. The results not only show that the stage of RNA isolation can b e bypassed but also illustrate an easy and efficient means of obtainin g templates suitable for identification and characterisation of HCV an d BVDV in tissue culture by RT-PCR.