| 西尼罗河病毒病竞争ELISA检测方法的初步建立及应用 |
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| 引用本文： | 常娓娓,王淑娟,吴晓东,刘华雷,赵永刚,王志亮.西尼罗河病毒病竞争ELISA检测方法的初步建立及应用[J].中国动物检疫,2011,28(9):31-34. |
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| 作者姓名： | 常娓娓 王淑娟 吴晓东 刘华雷 赵永刚 王志亮 |
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| 作者单位： | 中国动物卫生与流行病学中心国家外来动物疫病诊断中心,山东青岛,266032 |
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| 摘 要： | 本研究用表达的西尼罗河病毒(WNV)囊膜蛋白E结构域III蛋白作为包被抗原,单抗8F4A4作为竞争检测抗体,初步建立了能够对乙型脑炎病毒(JEV)和WNV进行鉴别诊断的竞争ELISA方法。优化的最佳抗原包被浓度为530ng/mL,单抗的最佳稀释倍数为1:8000,待检血清的最佳稀释倍数为1:10。通过对56份阴性样品进行检测确定了该方法的临界值为30%,以此为判定标准对临床220份血清进行检测,结果均为阴性。此方法的建立弥补了商品化试剂盒不能区分JEV和WNV感染的缺陷,为我国西尼罗河病毒病的流行病学调查提供了一种有效的抗体检测方法。
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| 关 键 词： | 西尼罗河病毒 囊膜蛋白 单克隆抗体 竞争ELISA |
Development and Application of a Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Antibody against West Nile Virus |
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| Chang Weiwei,Wang Shujuan,Wu Xiaodong,Liu Hualei,Zhao Yonggang,Wang Zhiliang.Development and Application of a Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Antibody against West Nile Virus[J].China Journal Of Animal Quarantine,2011,28(9):31-34. |
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| Authors: | Chang Weiwei Wang Shujuan Wu Xiaodong Liu Hualei Zhao Yonggang Wang Zhiliang |
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| Institution: | Chang Weiwei1,2,Wang Shujuan1,Wu Xiaodong1,Liu Hualei1,Zhao Yonggang1,Wang Zhiliang1(1.National Diagnostic Center of Animal Exotic Infectious Diseases,Chinese National Center for Animal Health and Epidemiology,Qingdao,266032,2.College of Veterinary Medicine,Yangzhou University,Yangzhou,225009) |
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| Abstract: | A cELISA for detection of WNV antibody was established using purified WNV E protein domain IH as antigen coating the ELISA plate and monoclonal antibody 8F4A4 specific to WNV as competitive detection antibody.The results showed that the antigen coat concentration was 530 ng/mL, the best dilution of McAb 8F4A4 was 1：8000 and the best serum dilution was I：10.A total of 56 clinically confirmed negative control sera were tested by the cELISA.Through analysis, it was concluded that the cut-off value was 30%.A set of 220 clinical serum samples were tested by the cELISA and all were negative.Contrast to the commercial kits, the method could differentiate the JEV and WNV antibody, providing an efficient antibody detection method for the West Nile virus disease epidemiology investigation in our country. |
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| Keywords: | West Nile virus （WNV） Envelope protein Monlclonal antibody cELISA |
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