结核分枝杆菌环介导恒温扩增(LAMP)快速检测方法的评价  

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

作　　者：戴广明[1] 曹以诚[2] 杜正平[2] 陈洵[2] 黄曙海 逄宇[1] 周杨[1] 黄海荣[1] 赵雁林[1] 

机构地区：[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 [3]广西疾病预防控制中心,南宁530000

出　　处：《中国防痨杂志》2011年第1期47-51,共5页Chinese Journal of Antituberculosis

基　　金：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

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 am-plification,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gyrB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60min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。Objective To evaluate the value of loop-mediated isothermal amplification（LAMP） in rapid detection of Mycobacterium tuberculosis.Methods A set of four LAMP primers were designed for recognizing six distinct regions on the target DNA（gyrB）.The DNAs of M.tuberculosis strains 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 diagnosing M.tuberculosis infection.

关 键 词：分枝杆菌 结核 核酸扩增技术 

分 类 号：R446.5[医药卫生—诊断学]