| PRRSV SCQ株M蛋白基因截断片段原核表达及间接ELISA检测方法的初步建立 |
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| 引用本文： | 陈进会,余霞,颜其贵,万莉,马磊,王志彪.PRRSV SCQ株M蛋白基因截断片段原核表达及间接ELISA检测方法的初步建立[J].检验检疫科学,2011(4):26-31. |
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| 作者姓名： | 陈进会 余霞 颜其贵 万莉 马磊 王志彪 |
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| 作者单位： | 东莞出入境检验检疫局;河源出入境检验检疫局;四川农业大学动物医学院; |
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| 基金项目： | 广东出入境检验检疫局科技项目(2010GDK64); 东莞市科技局项目(2007108101100); 四川省教育厅重点项目(07ZA053) |
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| 摘 要： | 通过PCR方法从重组质粒pMD18-T-M扩增得到缺失N端跨膜区的M蛋白基因片段dM（de-letingM）,克隆至高效原核表达载体pGEX-4T-1,得到重组M蛋白。以纯化的重组M蛋白作为抗原,建立检测猪繁殖与呼吸综合征病毒抗体的间接ELISA方法,并确立ELISA最佳工作条件。经重复性试验、交叉试验、阻断试验等试验验证,结果表明该方法重复性好、特异性强、敏感度高;与美国IDEXX公司试剂盒相比较,特异性和敏感性分别为96.3%和93.5%,无显著性差异。用建立的方法检测临床血清样品168份,总阳性率为39.9%。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 M蛋白基因 原核表达 间接ELISA |
Prokaryotic Expression of Truncated Fragment M Protein Gene and Establishment of Indirect ELISA Assay for Porcine Reproductive and Respiratory Syndrome Virus |
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| Chen Jinhui,Yu Xia,Yan Qigui,Wan Li,Ma Lei,Wang Zhibiao.Prokaryotic Expression of Truncated Fragment M Protein Gene and Establishment of Indirect ELISA Assay for Porcine Reproductive and Respiratory Syndrome Virus[J].Inspection and Quarantine Science,2011(4):26-31. |
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| Authors: | Chen Jinhui Yu Xia Yan Qigui Wan Li Ma Lei Wang Zhibiao |
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| Institution: | Chen Jinhui1,Yu Xia2,Yan Qigui3,Wan Li3,Ma Lei3,Wang Zhibiao3(1.Dongguan Entry-Exit Inspection and Quarantine Bureau,Dongguan,Guangdong,523072,2.Heyuan Entry-Exit Inspection and Quarantine Bureau,3.College of Veterinary Medicine,Sichuan Agricultural University) |
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| Abstract: | The gene segment dM(deleting M)deleting N transmembrane domain sequence was successfully amplified from recombinant plasmids pMD18-T-M by PCR.Then,the dM gene was cloned into plasmid of PGEX-4T-1 and recombinant protein M was expressed and harvested.Using purified matrix membrane protein,an indirect ELISA for detection of anti-PRRSV antibodies was developed and its optimal reaction conditions were determined.The ELISA assay was confirmed to have a good reproducibility,specificity and sensitivity by reproduc... |
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| Keywords: | PRRSV M Protein Gene Prokaryotic Expression Indirect ELISA |
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