Authors : J. R. Patel and R. W. Shilleto
Abstract: Procedures are described in Bovine Embryo Lung (BEL) cells for the detection and titration of Non-cytopathic Ruminant Pestiviruses (NCRPVs) and NCRPV neutralising (VN) antibodies based on the interference between NCRPVs and cytopathic Bovine Viral Diarrhoea Virus (BVDV). The interference based NCRPV detection and titration tests in BEL cells were highly reproducible and of equal sensitivity to the traditional immunofluorescence detection method in primary (<15 passages) bovine turbinate cells. The age of BEL cell monolayers did not affect the plating efficiency of NCRPVs. Thus in monolayers of 2, 4, 9 and 11 days of age, infectivity titres of six NCRPVs strains varied by 2-4 fold. The interference test in BEL cells is routinely used for the detection and titration of NCRPVs in cell lines, viral stocks, blood and swabs from cattle and sheep and viral antisera but more importantly in Foetal Calf Serum (FCS) which is the common source of infection of tissue culture by non-cytopathic BVDV strains. In spiked FCS the test could detect as little as 4 median Tissue Culture Infective Doses (TCID50) of NCRPV per 50 mls. The interference principle was also used to develop a routine test for titration of ruminant sera for VN antibody to NCRPV.
J. R. Patel and R. W. Shilleto , 2003. Ruminant Pestivirus Detection and Serology Based on Interference between Non-cytopathic and Cytopathic Viruses. Journal of Animal and Veterinary Advances, 2: 327-337.