小反刍兽疫病毒竞争ELISA检测试剂盒生产工艺研究

Research on the Production Process of Competitive ELISA Test Kit for Peste des Petits Ruminants Virus

为研究小反刍兽疫病毒(PPRV)竞争ELISA试剂盒的生产工艺,本试验利用昆虫细胞表达的PPRV核蛋白及其单克隆抗体,建立一种特异性高、敏感性强的PPRV竞争ELISA检测方法,并对该方法的各个步骤进行优化,确定该方法的最适条件,从而确定竞争ELISA的操作程序。并用该方法与OIE推荐的PPRV rC-ELISA抗体检测试剂盒同时检测来自西藏、内蒙古共977份临床羊、牛血清样本,结果显示特异性、敏感性、符合率分别达99.89%、93.82%、99.38%。本研究建立的PPRV竞争ELISA方法为该病毒检测试剂盒的研制奠定技术基础,为小反刍兽疫的防控提供科学依据。

In order to research on the production process of competitive ELISA test kit for peste des petits ruminants virus （PPRV）, in this test, nucleoprotein （N） protein expressed in insect cell and monoclonal antibody of PPRV were used to estab lish a high specificity, sensitivity and strong competitive ELISA method for detection of PPRV, and the competitive ELISA procedures were optimized. 977 sheep and cattle serum samples collected from Xizang and Inner Mongolia were assayed with this established competitive ELISA and rCELISA antibody kit for PPRV of the OIE at the same time. The detecting results displayed that specificity, sensitivity and accuracy were 99.89%, 93.82 % and 99.38%. The competitive ELISA method of PPRV established in this assay would provide the basis for research on the production process of ELISA test kit as well as a ref- erence for prevention scientifically.