Premilinary Study On Prevention And Control Technology Of Bluetongue Disease

Bluetongue (BT) is a non-contagious disease caused by bluetongue virus (BTV) which belongs to the Orbiviruse genus, the Reoviridae family. BTV can infect all ruminant species including sheep, cattle and other wild rumiants and the virus is an arbovirus which is transmitted between ruminant hosts by some species of the Culicoides genus. The average mortality of infected animals is30%, however, in sheep as high as80%. Up to now, BTV has at least26many serotypes around the world which could generate only low levels of cross-protection, and directly enhance the difficult of the disease control and prevention. Recently BTV8is outbreak in Europe, and avoked the research enthusiasm and consideration of the disease.In this study, recombinant VP2protein and BTV8inactivated virus were used to immunize BALB/c mice and successfully obtained three BTV8type-specific monoclonal antibody (MAbs)(2E7.2G9and3B7) and three BTV group-specific MAbs (1F6.4C2与6G12) by traditional cell fusion technology. And by fused pepetides expression and phage display technology confirmed two BTV8type-specific line epitopes and key amino acids which form the conformational epitopes recongnized by MAb4H7. Then the BTV group-specific MAb1F6and4H7were used to develop an antigen capture ELISA which can be used to quantify the BTV antigen, and has notable correlation compared to TCID50which is the effective way to determine titre of virus. At last, we got the purified BTV8by plaques cloning technology, and using hydroxylamine, β-propiolacton (BPL) and binary ethylenimine (BEI) as virus inactivating agents and ISA15VG and nano603as adjuvants, evaluate the most effective combination for BTV8inactivated vaccine. Meanwhile, by VP7competition antibody and sera neutralizing antibody (SNT) detection confirmed the optimal comnination is BPL virus inactivating agent with ISA15VG adjuvant.In the present study, we develope and identify of the biology characters of BTV8type-specific MAbs and BTV group-specific MAbs and used them to develop an antigen capture ELISA which can be used to quantify the BTV antigen. At last, we successful confirm the product process of BTV8inactivated vaccine which is important to BTV8prevention and control.