猪口蹄疫病毒VP1结构蛋白抗体间接ELISA方法的建立  

Establishment of Indirect ELISA Diagnose Based on the VP1 Structural Protein of Foot-and-mouth Disease Virus(FMDV)in Pigs

作　　者：王光华[1,2] 独军政[1] 丛国正[1] 邵军军[1] 林彤[1] 薛慧文[2] 常惠芸[1] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,国家口蹄疫参考实验室,兰州730046 [2]甘肃农业大学动物医学院,兰州730076

出　　处：《生物工程学报》2007年第5期961-966,共6页Chinese Journal of Biotechnology

基　　金：国家重点基础研究发展计划(No.2005CB523201);国家支撑计划(2006BAD06A03)~~

摘　　要：将口蹄疫病毒(FMDV)的VP1基因,通过pPROex-HT表达载体在大肠杆菌BL21(DE3)中成功表达,获得大小为31ku的融合蛋白,Westernblot检测证实表达的该蛋白具有良好的生物学活性。以纯化的融合蛋白为抗原建立了猪FMDVVP1蛋白间接ELISA检测方法。通过对80份田间血清样品的检测表明,该方法与FMDV液相阻断ELISA(国标试剂盒)的总符合率为96.25%,表明建立的VP1蛋白间接ELISA检测方法具有很好的特异性和敏感性。The complete gene encoding the structural protein of FMDV（VP1）was subcloned into expression vector pPROex-HT,resulting in the fusion expression plasmid pPROexHT-VP1.After transformed into E.coli BL21（DE3）and induced by IPTG,the fusion protein was expressed in high level.Western blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV.Based on the fusion protein further purified,a novel indirect ELISA（VP1-ELISA）was developed to detect FMDV antibody in pigs.Comparison between VP1-ELISA and the government standard kit（liquid phase block ELISA）showed the two methods had 96.25 percent agreement by detecting 80 serum samples,indicating that the indirect VP1-ELISA was specific and sensitive.

关 键 词：口蹄疫病毒抗体 VP1蛋白 间接ELISA 

分 类 号：S854.43[农业科学—临床兽医学] Q789[农业科学—兽医学]