标题：直接RT-LAMP检测EV71方法的建立和评价

      作者：阮美生 1，郭龙华 2，周天龙 1，黎 荣 1
    (1.湖北科技学院附属第二医院检验科，湖北 咸宁 437100；
2.深圳市龙华新区人民医院检验科，广东 深圳 518109）

      卷次： 2015年26卷18期

      【摘要】 目的 建立并评价一种简单快速的逆转录环-介导的等温扩增(直接RT-LAMP)检测人肠道病毒71
(EV71)的实验方法。方法 无需RNA提取，用热处理标本后用直接RT-LAMP方法检测EV71，并用临床收集
的 290份咽试标本评价其灵敏度和特异性。结果 直接 RT-LAMP和 RT-LAMP实验完全符合率为 92.4%
(268/290)，直接RT-LAMP和qRT-PCR完全符合率为88.9%。在RT-LAMP或直接RT-LAMP实验中没有发现假阳
性。与RT-LAMP比较，直接RT-LAMP的灵敏度和特异性分别为90.3%和100%；与qRT-PCR比较，其敏感性和特
异性分别为86.8%和100%。结论 直接RT-LAMP方法能被开发成简单快速检测EV71和其他病原体的方法。

      【关键词】 逆转录环-介导的同温扩增；肠道病毒71；实时定量RT-PCR；灵敏度；特异性

      【中图分类号】 R446 【文献标识码】 A 【文章编号】 1003—6350（2015）18—2717—04


Development and assessment of direct reverse transcription loop-mediated isothermal amplification for EV71
detection.
RUAN Mei-sheng 1, GUO Long-hua 2, ZHOU Tian-long 1, LI Rong 1. 1. Department of Clinical Laboratory, the
Second Affiliated Hospital, Hubei University of Science and Technology, Xianning 437100, Hubei, CHINA; 2. Department
of Clinical Laboratory, Shenzhen Longhua New District People's Hospital, Shenzhen 518109, Guangdong, CHINA

【Abstract】 Objective To develop a simple and rapid direct reverse transcription loop-mediated isothermal am-
plification (direct RT-LAMP) experimental method for detecting enterovirus 71 (EV71) infection, and the sensitivity and
specificity of this method using clinical nasopharyngeal swab specimens for the detection of EV71 were evaluated.
Methods Direct RT-LAMP without RNA extraction and with heat-treatment was used to detect EV71 infection in 290
nasopharyngeal swab specimens. The sensitivity and specificity were evaluated. Results The accordance rate of direct
RT-LAMP and RT-LAMP was 92.4% (268/290), and that of direct RT-LAMP and qRT-PCR was 88.9%. No false posi-
tive was found in direct RT-LAMP or RT-LAMP. The clinical performance demonstrated the sensitivity and specificity
of direct RT-LAMP was 90.3% and 100% compared to RT-LAMP, and 86.8% and 100% compared to qRT-PCR, re-
spectively. Conclusion Direct RT-LAMP method can potentially be developed for simple and rapid screening of
EV71 and other pathogens.

      【Key words】 Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Enterovirus 71