口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测方法的建立  

Development of time-resolved fluoroimmunoassay method for the detection of non-structural protein antibodies against foot-and-mouth disease virus

作　　者：孙雨[1] 宋晓晖[1] 肖颖 毕一鸣[1] 王睿男[1] 蒋菲[1] 张存瑞[1] 王旭 杨林[1] 王传彬[1] SUN Yu;SONG Xiao-hui;XIAO Ying;BI Yi-ming;WANG Rui-nan;JIANG Fei;ZHANG Cun-rui;WANG Xu;YANG Lin;WANG Chuan-bin(China Animal Disease Control Center,Beijing 102600,China;Chongqing Animal Disease Control Center,Chongqing 400000,China)

机构地区：[1]中国动物疫病预防控制中心,北京102600 [2]重庆动物疫病预防控制中心,重庆400000

出　　处：《中国兽医科学》2019年第7期812-820,共9页Chinese Veterinary Science

基　　金：国家重点计划研发项目(2016YFD0501504)

摘　　要：为建立快速定量检测羊血清中口蹄疫病毒非结构蛋白抗体的方法,本研究通过优化抗原表达条件等步骤,在大肠杆菌原核表达系统中获得了可溶性的3A-3B1-3B2融合蛋白,基于纯化的可溶性融合蛋白建立了口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测试剂盒。该方法能够检测羊血清中的口蹄疫病毒非结构蛋白抗体,敏感性高,特异性强,对其他相关的羊类病原无交叉反应,其组内与组间变异系数分别低于10%和15%,具有良好的重复性。对300份临床血清样品进行检测,同Procheck公司的口蹄疫非结构蛋白抗体试剂盒进行比较,阳性样品符合率为98%,阴性样品符合率为92%,总符合率为95%。重复性试验组内与组间变异系数均小于10%。口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测方法同传统的ELISA方法相比,特异性相当、敏感性更高,操作简单、快速,具有较高的应用推广价值。To establish the time-resolved fluoroimmunoassay method for the detection of foot-andmouth disease virus antibodies in goat serum,this research obtained the soluble 3A-3B1-3B2 fusion protein in Escherichia coli,through exploration of protein expression conditions.Further based on the purified fusion protein,a time-resolved fluoroimmunoassay detection method for foot-and-mouth disease virus antibodies was established.This method could detect the antibodies against foot-and-mouth disease virus in goat serum quickly and quantitatively.It had high sensitivity and specificity,and had no cross reaction to other related goat pathogens.The coefficient of variation intra batch and inter batch were all below 10%.A total of 300 clinical serum samples were tested,and the ELISA kit(Procheck)was used for comparison.The coincidence rate of the positive samples was 98%,the coincidence rate of the negative samples was 92%,and the total coincidence rate was 95%.In conclusion this method has a wide range of detection with high specificity,and has a high practical value and popularization value,compared with the traditional ELISA.

关 键 词：口蹄疫病毒 非结构蛋白 可溶性表达与纯化 时间分辨荧光免疫分析 

分 类 号：S852.659.6[农业科学—基础兽医学]