Establishment And Application Of Competitive ELISA For Detection Of Antibody Against Bluetongue Virus

Bluetongue virus(BTV)causes Bluetongue(BT)in domestic or wild ruminants.The disease is transmitted by insect vectors such as cumidge and is a typical non-contact viral infection.At present,this disease is one of the class A diseases defined by OIE,and our country has defined it as A class of animal infectious diseases.The bluetongue virus belongs to reoviridae,orbivirusand the BTV particle contains a three-dimensional symmetry of 20 planes,round,without capsule,and a diameter of about 53~60 nm,presenting a hollow short cylindrical shape.The core is made up of 10 linear double-stranded RNA.BTV housing is mainly composed of VP2 and VP5.The inner shell of the virus is mainly composed of VP3 and VP7.VP7 in the nucleic capsid existsmainly in the form of trimer,and is a BTV group specific antigen.BT has 28 serotypes,and these serotypes have no cross-protective effect.It is imperative that a groupspecific blocking effect of ELISA detection kits by independent research and development be prepared,for our country’s epidemiological investigation and monitoring provides a quick,safe and low cost technology.After IPTG induced expression of peT28a-8S7/BL21,the protein was purified by resin,and the results that the protein can produce specific reaction with bluetongue virus 24 standard positive serum,suggests that the VP7 protein has a good group specific reactivity by the expression of the 8 type blue tongue virus.In this study,VP7 protein was used as antigen,Anti-8 bluetongue virus of VP7 proteina of monoclonal antibody 6D11 as a competitive antibody,and a method was established.The method was compared with the ELISA kit produced by the company for detection of bluetongue antibody,and 172 serum samples of goats,sheep and cattle in different regions were detected,and the coincidence rate of sample test results was as high as 98.84%.The method can be used for epidemiological investigation and monitoring,and the establishment of this method provides safe,fast and accurate technical means for the detection of serology in China.