Zastosowanie reakcji polimeryzacji lancuchowej w wykrywaniu zakazen wirusami z rodzaju Pestivirus

Reverse transcription coupled with the polymerase chain reaction (RT-PCR) was used for the detection of HCV and BVDV in virus cell cultures. Reverse transcription reaction was performed with RNA isolated by means of the guanidine method or with cell culture supernatant precipitates by eliminating the RNA isolation step. Both reactions, RT and PCR, were carried out by using rTth DNA polymerase in one tube. The RT-PCR products were analysed by a gel electrophoresis and hybridization with the digoxygenine labelled DNA probe. It was demonstrated that it is possible to omit the RNA isolation step and that applied primers are suitable for diagnosing HCV and BVDV.