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小反刍兽疫病毒(PPRV)N蛋白是病毒粒子的主要结构蛋白,也是诊断小反刍兽疫的主要靶蛋白。为获得高效表达的可溶性N蛋白,并建立基于N蛋白的小反刍兽疫的间接ELISA检测方法,通过优化密码子、优化表达条件等条件探索,在大肠埃希菌表达系统中获得了高效表达的可溶性N蛋白,进一步基于N蛋白建立了间接ELISA检测方法,该方法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均低于9%,具有良好的重复性。对480份临床血清样品进行检测,同时用法国IDVET竞争ELISA试剂盒进行比较,符合率达到98.33%。本研究为进一步开发成熟的小反刍兽疫抗体检测试剂盒奠定了基础。
The PPRV N protein is the major structural protein of virus particles and is also the main target protein for the diagnosis of PRRSV. In order to obtain highly expressed soluble N protein and to establish an indirect ELISA method based on N protein for the determination of PPR, we optimized the codon usage and optimized the expression conditions to obtain the highly expressed soluble protein in Escherichia coli expression system N protein. The indirect ELISA method was established based on the N protein. The method has no cross-reaction to other related pathogenicity of sheep. The coefficient of variation (CV) within and between the two groups are all less than 9%, with good repeatability. 480 clinical serum samples were tested, and compared with France IDVET competition ELISA kit, the coincidence rate reached 98.33%. This study lays the foundation for the further development of a developed kit for the detection of antibodies to PRRSV.