鉴别蓝舌病野毒感染ELISA检测方法的建立与初步应用  

Development of diagnostic kit of infected and immuned animals of BTV

作　　者：黄超华 史卫军 曹琛福 王潇 花群义 林彦星 陈金顶[1] HUANG Chao-hua;SHI Wei-jun;CAO Chen-fu;WANG Xiao;HUA Qun-yi;LIN Yan-xing;CHEN Jin-ding(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Animal and Plant Inspection and Quarantine Technology Center,Shenzhen Customs District of P.R.C.,Shenzhen,Guangdong 518045,China)

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]深圳海关动植物检验检疫技术中心,广东深圳518045

出　　处：《中国兽医学报》2020年第8期1438-1442,共5页Chinese Journal of Veterinary Science

基　　金：国家“十三五”重点研发计划资助项目(2017YFD0502304,2017YFD0501104);广州市科技计划资助项目(201803020005)。

摘　　要：利用原核表达系统表达并纯化获得蓝舌病病毒NS3蛋白。以NS3重组蛋白作为包被抗原,经过对包被浓度、样品稀释度以及二抗工作浓度和作用时间等一系列条件的优化,建立了一种鉴别蓝舌病野毒感染的间接ELISA抗体检测方法。结果显示,该方法的检测灵敏度可达1∶1280,批内、批间变异系数均小于10%,具有良好的重复性。与商业化的蓝舌病试剂盒对比检测的结果显示,两者的蓝舌病阳性样品符合率为100%,本试验建立的间接ELISA方法可以区分蓝舌病自然感染和疫苗免疫动物。In order to develop an assay to differentiate diagnosis the infected and immuned animals of bluetongue virus(BTV),this study applied the prokaryotic expression system to express BTV NS3 protein.After optimization of the reaction conditions,an indirect ELISA for the detection of the antibody in sera was developed by using the recombinant protein NS3 as a coating antigen.The result showed that the detection limit of the indirect ELISA could reach 1∶1280 dilution of BTV standard positive serum,and the variation of inter-batch and intra-batch were less than 10%.Compared with the commercial BTV ELISA kit,it was confirmed that the developed indirect ELISA could distinguish the immuned animals from infected animals of BTV.

关 键 词：蓝舌病病毒 NS3蛋白 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]