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

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，Western blot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FMDV VP1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。

Abstract:

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.