Email Alert | RSS    帮助

中国防痨杂志 ›› 2011, Vol. 33 ›› Issue (1): 47-51.

• 论著 • 上一篇    下一篇

结核分枝杆菌环介导恒温扩增(LAMP)快速检测方法的评价

戴广明1, 曹以诚2, 杜正平2, 陈洵2, 黄曙海3, 逄宇1, 周杨1, 黄海荣1, 赵雁林1   

  1. 1.北京结核病胸部肿瘤研究所 北京 101149; 2.华南理工大学 广州 510640; 广西疾病预防控制中心 南宁 530000;
  • 收稿日期:2010-01-04 出版日期:2011-01-20 发布日期:2012-01-19
  • 通信作者: 赵雁林 E-mail:zhaoyanlin@tb123.org
  • 基金资助:

    北京市科技新星计划(2007B039); 中-法梅里埃基金会结核病防治项目

Evaluation of loop-mediated isothermal amplification (LAMP) in rapid detection of Mycobacterium tuberculosis

Dai Guangming,Cao Yicheng,Chen Xun,Du Zhengping,Huang Shuhai,Pang Yu,Zhou Yang,Huang Hairong,Zhao Yanlin   

  1. 1.Beijing chest hospital, Beijing 101149, China; 2.South China University of Technology, Guangzhou 510640, China; 3.CDC of Guangxi province, Nanjing 530000, China
  • Received:2010-01-04 Online:2011-01-20 Published:2012-01-19
  • Contact: Zhao Yanlin E-mail:zhaoyanlin@tb123.org

摘要: 目的 对国内建立的结核分枝杆菌DNA环介导恒温扩增(loop-mediated isothermal amplification, LAMP)快速检测方法进行了介绍和评价。 方法 实验菌株来源于北京结核病胸部肿瘤研究所。以结核分枝杆菌作为研究对象,设计了4 条LAMP 引物特异地识别结核分枝杆菌中 gyr B 基因的6个特殊区域。所有的菌株样本提取DNA作为模板,加入LAMP反应管后,65 ℃恒温反应60 min即可。其结果可以通过肉眼观察到的颜色变化来判断。 结果 实验共验证了79株临床和标准菌株。对于菌株的培养物,LAMP实验的灵敏度为100%,特异度为92%。 结论 结核分枝杆菌LAMP快速检测方法是一种快速、可靠的结核分枝杆菌诊断方法。

关键词: 分枝杆菌, 结核, 核酸扩增技术

Abstract: Objective To evaluate the value of loop-mediated isothermal amplification (LAMP) in rapid detection of Mycobacterium tuberculosisMethods A set of four LAMP primers were designed for recognizing six distinct regions on the target DNA (gyrB). The DNAs ofM. tuberculosisstrains from Beijing Chest Hospital were extracted,amplified at 65°C for 60 min, and their reaction Results were observed by simple naked eye.   Results A total of 79 clinical isolates and standard strains were studied. The sensitivity and specificity of LAMP was 100% and 92%, respectively.  Conclusions LAMP is a rapid and reliable alternative method for diagnosingM. tuberculosisinfection.

Key words: M. tuberculosis, nucleic acid amplification techniques