摘要
克隆、表达了猪繁殖与呼吸综合征病毒DY株(GenBank:JN864948)的Nsp4蛋白,并利用Nsp4作为包被蛋白建立了ELISA检测方法。优化的Nsp4抗体ELISA检测方法的最佳抗原包被浓度为1μg/mL,血清最佳稀释度为1∶40,最佳封闭液为100mL/L牛血清,酶标二抗的最佳工作稀释度为1∶4000,最佳显色时间为37℃反应15min。用建立的ELISA检测方法和IDEXX ELISA检测试剂盒检测血清样品130份,总符合率为93.19%。Nsp4抗体ELISA检测方法的建立,可为PRRSV的抗体监测提供技术支持。
In this study,Nsp4 gene was cloned and expressed from porcine reproductive and respiratory syndrome virus DY strain (GenBank:JN864948),and a Nsp4 antibody ELISA detection method was established by using Nsp4 protein.The optimal antigen package was 1 μg/mL,the best dilution of the serum was 1∶40,the best closed liquid was 100 ml/L bovine serum,the best working dilution of enzyme labeled secondary antibody was 1∶4 000,and the best coloring time at 37℃ was 15 min.For compliance experiment,130 serum samples were detected by IDEXX ELISA Kit,and the total compliance rate was 93.19%.The establishment of Nsp4 antibody ELISA detection method provides a new idea for PRRSV antibody monitoring.
作者
刘磊
马永彪
王文秀
李峰
沈志强
张松林
LIU Lei;MA Yong-biao;WANG Wen-xiu;LI Feng;SHEN Zhi-qiang;ZHANG Song-lin(Shandong Binzhou Animal Science&Veterinary Medicine Academy,Binzhou,Shandong,256600,China;Shandong Lvdu Bio-sciences &technology Co.,LTD,Binzhou,Shandong,256600,China)
出处
《动物医学进展》
北大核心
2019年第7期9-13,共5页
Progress In Veterinary Medicine
基金
山东省重点研发计划项目(2016GNC110032)
山东省自然科学基金项目(ZR2017PC003)
山东省现代农业产业技术体系生猪创新团队项目(SDAIT-08-17)
关键词
猪繁殖与呼吸综合征病毒
非结构蛋白
抗体检测
Porcine reproductive and respiratory syndrome virus
non-structural protein
antibody detection
