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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

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【作者】 戴广明; 曹以诚; 杜正平; 陈洵; 黄曙海; 逄宇; 周杨; 黄海荣; 赵雁林;

【Author】 Dai Guangming1,Cao Yicheng1,Chen Xun2,Du Zhengping2,Huang Shuhai3,Pang Yu1, Zhou Yang1,Huang Hairong1,Zhao Yanlin1(1.Beijing chest hospital,Beijing 101149,China; 2.South China University of Technology,Guangzhou 510640,China; 3.CDC of Guangxi province,Nanjing 530000,China)

【机构】 北京结核病胸部肿瘤研究所; 华南理工大学; 广西疾病预防控制中心;

【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。

【Abstract】 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.

【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
  • 【文献出处】 中国防痨杂志 ,Chinese Journal of Antituberculosis , 编辑部邮箱 ,2011年01期
  • 【分类号】R446.5
  • 【被引频次】16
  • 【下载频次】404
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