Establishment Of Indirect ELISA Assay For Bovine Viral Diarrhea/Mucosal Virus And Its Serological Investigation

Bovine viral diarrhea virus (BVDV) is one member of the Flaviviridae, Pestivirus. It’s a seriouspathogen seriously endanger to the livestock’ health around the worldwide. This disease can cause patienthigh fever，oral and nasal mucosal bleeding or mucus oversecretion，reduction of milk production even offproduction，pregnant cow abortion or stillbirth，featus anomalies and other symptoms. BVDV hasserological cross-reactions with CSFV and BDV. So far,there is no effective prevention and controlmeasures of BVDV.Therefore,to establish a rapid and efficient detection method for early detection ofBVDV is very important for early prevention.In this study, after being passaged in MDBK cells, BVDV was purified by ultracentrifugation, andan indirect enzyme-linked immunosorbent assay(ELISA) for detecting serum antibody against BVD wasdeveloped and optimalized. After being coated with BVDV which was diluted with phosphate buffer to1:200at4℃for12h, the ELISA plates were blocked with5%nonfat dried milk at37℃for2h, incubatedwith test sera diluted1:100at37℃for1h, and followed by an incubation with rabbit anti-bovine IgGantibody conjugated with HRP at a dilution of1:10000at37℃for30min. The cut off values are0.320.The results showed that the established indirect ELISA had high specificity and sensitivity, smallvariability. The agreement of the indirect ELISA with the commercialization of BVD antibody detection kitconformity was91.3%.In summary, the established indirect ELISA for detecting serum antibody against BVD providedpowerful support for monitoring and controlling BVD in Xinjiang.