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基于RT-LAMP的10种烈性病毒快速检测方法的研究

  • 赵娜1 ,
  • 赵望2 ,
  • 刘淇3 ,
  • 夏菡1 ,
  • 师永霞4 ,
  • 袁志明1
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  • 1. 中国科学院武汉病毒研究所,武汉 430071;上海科源电子科技有限公司,上海 201611;复旦大学环境科学与工程系,上海 200433;4. 广东出入境检验检疫局检验检疫技术中心,广州 510627

收稿日期: 2018-01-04

  网络出版日期: 2018-10-25

基金资助

科技基础性工作专项(2013FY113500),广东省科技计划项目(2015A020213007)

Development of a method for rapid detection of 10 high-pathogenic viruses based on reverse transcription-loop-mediated isothermal amplification

  • ZHAO Na1 ,
  • ZHAO Wang2 ,
  • LIU Qi3 ,
  • XIA Han1 ,
  • SHI Yongxia4 ,
  • YUAN Zhiming1
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  • 1.Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China; 2.Shanghai Cohere Electronics Technology Co. Ltd, Shanghai 201611, China; 3.Department of Environmental Science & Engineering,Fudan University, Shanghai 200433, China; 4.Guangzhou Inspection & Quarantine Technology Center, Guangzhou 510627, China

Received date: 2018-01-04

  Online published: 2018-10-25

摘要

本研究针对10种对人类健康威胁较大的烈性病毒建立反转录-环介导等温扩增法(reverse transcription-loop-mediated isothermal amplification,RT-LAMP),筛选出10种烈性病毒高特异性引物,并分别对这10种病毒进行特异检测。结果显示,其安全、便捷、快速,且高通量。在自主研发的RT-LAMP体系中,只需加入模板,结合配套仪器,扩增耗时不超过45 min,检测灵敏度为1~100拷贝/反应(25 μL)。检测30份人血清样品,筛查出1份裂谷热病毒阳性。该方法操作简单,可防止污染,成本低,易携带,方便于低配实验室或现场进行实时高通量检测。结果提示,针对10种烈性病毒的RT-LAMP方法可对口岸等人员流动性大的地区疑似患者进行快速筛查,展现出良好的应用前景。

本文引用格式

赵娜1 , 赵望2 , 刘淇3 , 夏菡1 , 师永霞4 , 袁志明1 . 基于RT-LAMP的10种烈性病毒快速检测方法的研究[J]. 微生物与感染, 2018 , 13(5) : 265 -272 . DOI: 10.3969/j.issn.1673-6184.2018.05.002

Abstract

The present study aims to establish a method for rapid detection of a panel of pathogenic viruses based on reverse transcription-loop-mediated isothermal amplification (RT-LAMP). Primer sets of 10 pathogenic viruses with high specificity were screened. It was suggested that the method was safe, rapid, convenient, and high-throughput in detection of pathogenic viruses. When combined with the new immobilized reaction tubes, the amplification for 10 pathogenic viruses in 1-100 copies could be finished within 45 min. In the assay of 30 serum samples, one rift valley fever virus-positive sample was obtained. A rapid, convenient, low-cost, real-time and high-throughput detection method for common pathogenic viruses was established. The results indicate that RT-LAMP method for 10 high-pathogenic viruses has a good application prospect in rapidly screening suspected patients in areas with high population mobility such as ports.

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