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口蹄疫病毒3AB克隆表达及其抗体间接ELISA检测方法建立与应用研究

Study on Indirect ELISA Technique for the Detection of 3AB Antibody of Foot-and-Mouth Disease Virus (FMDV) and Cloning and Expression of 3AB Gene of FMDV
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摘要 采用重叠PCR方法人工合成3AB基因片段,将其克隆到PET-28a+载体,转化导入宿主菌BL21(DE3)中成功表达,获得大小约为33Ku的重组表达蛋白。以纯化的重组表达蛋白为抗原,建立了检测猪FMDV 3AB NSP抗体的间接ELISA方法。将本方法与进口口蹄疫非结构蛋白3ABC阻断ELISA试剂盒进行比较,结果显示,本方法可以用于区分猪口蹄疫自然感染动物与疫苗免疫动物。 The 3AB gene was assembled by overlaping PCR, and cloned into vector PET- 28a ＋, After transformed into E. coli BL21 （ DE3 ） and induced by IPTG, the target protein was expressed successfully. A FMDV - NSP 3AB indirect ELISA detection method was developed by using the purified recombinant non - structural protein 3AB as detected antigen. Compare with this established method and the kit from PRIONICS company, the result showed that this method can be used for differentiating FMDV infected from vaccinated animals.
作者 朱文钏 孔繁德 林祥梅 徐淑菲 吴德峰 韩雪清 吴绍强
机构地区 厦门出入境检验检疫局 福建农林大学动物科学学院 中国检验检疫科学研究院
出处 《检验检疫学刊》 2010年第6期4-8,38,共6页 Journal of Inspection and Quarantine
基金 国家质检总局科研项目(2007IK039)
关键词 猪 口蹄疫 非结构蛋白3AB ELISA 检测方法 Pig FMDV NSP-3AB ELISA Detection Method
分类号 S855.3 [农业科学—临床兽医学] R446.61 [医药卫生—诊断学]
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检验检疫学刊
2010年 第6期

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