Detection Of Bovine Viral Diarrhoea Virus. By Nucleic Acid Hybridization

Abstract

This investigation involved the use of the p80 gene of bovine viral diarrhoea virus (BVDV) as a probe for detection of BVDV by in situ and slot blot hybridization. The probe was labelled with digoxigenin and reactions were detected with antidigoxigenin alkaline phosphatase conjugate and nitroblue tetrazolium chloride and 5- bromo-4-chloro-3-indolyl phosphate. Negative results were occasionally obtained when in situ hybridization was used for detection of the cytopathic, NADL strain of BVDV cultured in Madin Darby bovine kidney (MDBK) cells. The probe was used in slot blot hybridization tests with noncytopathic field isolates of BVDV, passaged once in MDBK cells. Of twenty samples positive for noncytopathic BVDV by fluorescent antibody staining, seventeen were detected by slot blot hybridization. Based on these results, in situ and slot blot hybridization do not appear to be applicable as routine diagnostic tests for BVDV.