Detection of bovine viral diarrhea virus in infected cells by polymerase chain reaction [PCR]

ABSTRACT

Polymerase chain reaction [PCR] was used to detect bovine viral diarrhea [BVD] virus in infected bovine turbinate [BT] cell culture. Viral RNA was extracted, reverse transcribed to cDNA, and a specific fragment was amplified by PCR using specific primers. BVD virus specific probe was prepared by labeling plasmid DNA contained BVD, PBV4-P80 insert with 32p, using the nick translation technique. The amplified cDNA after agarose gel electrophoresis was transferred to nirocellulose paper by the Southern Blot technique and then hybridized with the probe. The reaction was visualized by autoradiography. It appeared that the technique is highly sensitive for detection of BVD virus in infected cells and could be used for rapid and accurate diagnosis