| 环介导恒温扩增技术快速检测非洲猪瘟病毒 |
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| 引用本文： | 王彩霞,刘建,林祥梅,吴绍强.环介导恒温扩增技术快速检测非洲猪瘟病毒[J].动物医学进展,2010,31(2):15-19. |
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| 作者姓名： | 王彩霞 刘建 林祥梅 吴绍强 |
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| 作者单位： | 中国检验检疫科学研究院动植物检疫研究所,北京,100029 |
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| 基金项目： | 国家科技支撑计划项目(2006BAD06A14) |
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| 摘 要： | 以人工合成的非洲猪瘟病毒(ASFV)P72全长基因为模板,利用在线软件Primer Explorer4设计环介导恒温扩增(LAMP)的内、外引物,通过对反应体系和反应条件进行优化,建立了ASFV的LAMP检测方法,同时进行了敏感性和特异性试验。结果表明,所建立的LAMP检测方法最低检测限可达10拷贝质粒DNA,且具有良好的特异性,LAMP产物的酶切鉴定也验证了反应的特异性。对凝胶电泳后紫外观察、肉眼观察颜色变化、生成沉淀观察以及实时荧光PCR扩增曲线Ct值判定等不同判定方法进行比较表明,以上结果判定方法各有其优缺点,其中荧光PCR的敏感性最高,染色法相对较方便直观,电泳法敏感性稍低。
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| 关 键 词： | 非洲猪瘟病毒 环介导恒温扩增 敏感性 特畀性 |
Detection of African Swine Fever Virus (ASFV) by Loop-mediated Isothermal Amplification Method |
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| WANG Cai-xia,LIU Jian,LIN Xiang-mei,WU Shao-qiang.Detection of African Swine Fever Virus (ASFV) by Loop-mediated Isothermal Amplification Method[J].Progress In Veterinary Medicine,2010,31(2):15-19. |
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| Authors: | WANG Cai-xia LIU Jian LIN Xiang-mei WU Shao-qiang |
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| Institution: | Institute of Animal and Plant Quarantine;Chinese Academy of Inspection and Quarantine;Beijing;100029;China |
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| Abstract: | In order to establish a rapid method for detection African swine fever virus (ASFV),a loop-mediated isothermal amplification (LAMP) method was selected and the primers were designed using software Primer Explorer V4. The LAMP reaction solution and condition were optimized using the artificially synthesized ASFV antigenic gene of p72 as DNA template. The specificity and sensitivity of the developed ASF detecting LAMP method were examined. Results showed that the detected limit of ASFV detecting LAMP method w... |
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| Keywords: | African swine fever virus loop-mediated isothermal amplification sensitivity specificity |
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