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

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

Molecular differentiation of bovine viral diarrhea virus strains by radioimmunoprecipitation of viral polypeptides

Polypeptides of the reference cytopathic strain NADL, and the non- cytopathic strain NY-1 of bovine viral diarrhea [BVD] virus as well as another five non-cytopathic isolates of the same virus were examined by radioimmunoprecipitation of infected bovine fetal spleen by BFS cells. Viral proteins were analyzed by sodium dodecylsulfate- polyacrylamide gel [SDS-PAGE] electrophoresis. Polypeptides of molecular weight of 170 kDa, 130 kDa, 80 kDa, 60 kDa, 539 and 50 kDa were prominent for the NADL strain. The non-cytopathic NY-1 strain showed the same polypeptides profile as the NADL strain except that the polypeptide of 80 kDa was absent. The other five non-cytopathic strains showed the same polypeptides except in two strains where they have an additional two polypeptides of 56 and 54 kDa. But the 50 kDa polypeptide was absent. These results are of diagnostic and epidemiological significance