| 小反刍兽疫病毒N蛋白原核表达与间接ELISA检测方法的建立 |
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| 引用本文： | 孙雨,赵柏林,王晓英,董浩,翟新验,曲萍,胡冬梅,杨天意,石慧.小反刍兽疫病毒N蛋白原核表达与间接ELISA检测方法的建立[J].动物医学进展,2017,38(1). |
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| 作者姓名： | 孙雨 赵柏林 王晓英 董浩 翟新验 曲萍 胡冬梅 杨天意 石慧 |
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| 作者单位： | 中国动物疫病预防控制中心,北京,102600 |
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| 摘 要： | 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。
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| 关 键 词： | 小反刍兽疫 N活性蛋白 可溶性表达与纯化 间接酶联免疫吸附试验 |
Prokaryotic Expression of N Protein of Peste des Petits Ruminants Virus and Development of Indirect ELISA |
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| Abstract: | Peste des petits ruminants virus (PPRV) N protein is the major structural protein of virus particles,and it is the main target protein in the diagnosis of PPRV.To establish the expression and purification methods for efficient soluble PPRV N protein and indirect ELISA method for diagnosis of PPRV based on N,this study got a high expression of soluble N protein in Escherichia coli expression system through optimization of codon and expression conditions,and established an indirect ELISA detection method based on N protein.The results showed that the assay was specific and repeatable.Furthermore,a total of 480 clinical serum samples were detected by this assay and the agreement was 98.33% with commercial ELISA Kit (IDVET).This study laid the foundation for the further development of kit for detection of PPRV antibody. |
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| Keywords: | Peste des petits ruminants N active protein soluble expression and purification indirect ELISA |
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