猪副流感病毒5型HN蛋白的原核表达及间接ELISA方法的建立  

Prokaryotic Expression of Porcine Parainfluenza Virus 5 Hemagglutinin-neuraminidase and Establishment of Indirect ELISA Method

作　　者：倪民婷 张耕馨 孙普[2] 刘光亮[2] 陈佳宁 王金涛 NI Min-ting;ZHANG Geng-xin;SUN Pu;LIU Guang-liang;Chen Jia-ning;WANG Jin-tao(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang,163319,China;State Key Laboratory for Animal Disease Control and Prevention/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu,730046,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室,甘肃兰州730046

出　　处：《动物医学进展》2025年第3期52-57,共6页Progress In Veterinary Medicine

基　　金：黑龙江省重点研发项目(GA21B004);黑龙江八一农垦大学引进人才科研启动计划(XYB201913);黑龙江省生猪现代农业产业技术协同创新推广体系;国家生猪技术创新中心牧原项目(NCTIP-MY23004)。

摘　　要：为建立5型猪副流感病毒(Porcine parainfluenza virus 5,PPIV5)的血清学检测方法,用原核表达系统对PPIV5 HN蛋白进行重组表达,并用该重组蛋白进行PPIV5 ELISA检测方法的建立。结果显示,重组HN蛋白大小约为60 ku,反应性良好,以200 ng/孔包被抗原,10%BSA封闭30 min,1∶400稀释待检血清,1∶10000稀释二抗孵育30 min,显色5 min为最佳反应条件,该方法与猪圆环病毒、猪流行性腹泻病毒等阳性血清均无交叉反应,特异性好,PPIV5阳性血清最低检测稀释度为1∶800,敏感性高,批内和批间的变异系数均小于5%,重复性好。表明建立的ELISA方法具有良好的特异性、敏感性和重复性,为PPIV5的血清学诊断奠定了基础。To establish a serological method of porcine parainfluenza virus 5(PPIV5).The prokaryotic expression technique was used to reconstruct PPIV5 HN protein,and the recombinant protein was used to establish the PPIV5 ELISA method.The results showed that the size of recombinant HN protein was about 60 ku,and its reactivity was good.The 200 ng coated antigen,10%BSA closure for 30 min,1∶400 diluted serum,1∶10000 diluted secondary antibody for 30 min,and color development for 5 min were the optimal conditions.This method had no cross-reaction with porcine circovirus,porcine epidemic diarrhea virus and other positive sera.The minimum dilution of PPIV5 positive sera was 1∶800.The coefficient of variation and interbatches was less than 5%.The above results indicated that the ELISA method established in this study had good specificity,sensitivity and repeatability,which laid a foundation for serological diagnosis of PPIV5.

关 键 词：猪副流感病毒5型 HN蛋白 原核表达 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]