新型鸭呼肠孤病毒σC蛋白间接ELISA检测方法的建立  

Development of an indirect ELISA for detecting specific antibody to σC protein of Novel duck reovirus

作　　者：王锦祥[1,2] 程晓霞[1,2] 陈少莺[1,2] 陈仕龙[1,2] 林锋强[1,2] 王劭[1,2] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350013 [2]福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《中国预防兽医学报》2015年第9期687-690,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863项目(2011AA10A209);福建省自然科学基金项目(2014J05036)

摘　　要：为建立快速检测新型鸭呼肠孤病毒(NDRV)抗体的方法,本研究以纯化的重组σC蛋白作为包被抗原,并对该方法的反应条件进行优化,建立了NDRV间接ELISA抗体检测方法。该方法仅对NDRV血清检测为阳性,与番鸭呼肠孤病毒、鸭肝炎病毒、番鸭细小病毒、番鸭源鹅细小病毒阳性血清均无交叉反应,具有良好的特异性。其批内和批间重复性试验的变异系数均小于5%,具有良好的重复性。利用建立的σC蛋白间接ELISA方法对80份疑似鸭血清样品进行检测,结果显示与NDRV全病毒间接ELISA的符合率为88.75%。本研究建立的ELISA方法为NDRV的流行病学调查提供了快速、特异的血清学检测方法。To establish an assay for detection of the antibody against Novel duck reovirus （NDRV）, an indirect ELISA was developed with the purified recombinant σC protein as coating antigen expressed in E.coli. The optimal reaction conditions were determined as follow： coating with 625 ng of the σC protein per well for the microplate, blocking with 12% FBS buffer, diluting the serum sample at 1：80 and detecting with goat anti-duck HRP-IgG at 1：400. This method was specific for NDRV detection, and had no cross-reactions with sera against other duck viruses. The coefficient of intra- and inter-assay variations was both less than 5%. Moreover, the coincidence of the indirect ELISA coated with recombinant σC protein was 88.75% with the ELISA coated with the NDRV. The results indicated that the indirect σC-ELISA with sensitivity, specificity and repeatability would provide a rapid assay for detection of NDRV infection and serosurvey of the disease.

关 键 词：新型鸭呼肠孤病毒 原核表达 重组σC蛋白 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]