环介导等温扩增法快速检测结核杆菌复合群内主要致病菌  

Rapid Detection of the Main Pathogens of Mycobacterium Tuberculosis Complex by Loop-mediated Isothermal Amplification Assay

作　　者：洪明[1] 查磊[1] 付文亮[1] 邹民吉[1] 李伍举[1] 吴奎武[1] 徐东刚[1] 

机构地区：[1]中国人民解放军军事医学科学院基础医学研究所基因工程研究室,北京100850

出　　处：《医学研究杂志》2010年第11期60-63,共4页Journal of Medical Research

摘　　要：目的快速诊断和区分结核杆菌复合群内两种主要致病菌。方法利用环介导等温扩增(LAMP)技术建立快速检测和区分结核杆菌复合群内主要致病菌的方法,利用该方法对相关的临床分离株样本进行特异性检测,并利用1∶10倍比稀释的已知菌株DNA模板分析其敏感性。反应结束后通过电泳或向反应管中加入DNA染料肉眼判定检测结果。结果该方法可以成功检测到主要的结核致病菌:人型和牛型结核分枝杆菌,与包括卡介苗在内的其余相关菌株均未见非特异性交叉反应。检测的灵敏度可达100拷贝/微升,高于常规PCR方法。结论该检测方法具有敏感、特异、低成本和快速的特点,可检测和区分人型和牛型结核分枝杆菌,并能排除卡介苗对诊断的干扰。Objective To detecte two kinds of main pathogens of Mycobac - terium Tuberculosis complex rapidly. Methods We detected twelve kinds of bacteria samples by Loop- Mediated Isothermal Amplification（LAMP） and validated the sensitivity of LAMP by serial diluted DNA templates. The results were judged with naked eyes and electrophoretic analysis. Results The positive signal was only observed in Mtuberculosis vat. hominis and M. tuberculosis var. bovis, and no positive signal was observed for the other bacteria in the test. The detection limit of the assay was lcopy/μl. Conclusion This LAMP assay has many advantages such as specificity, sensitivity and simplicity. It is useful for rapid detection of tuberculosis, especially in avoiding the interference of BCG.

关 键 词：环介导等温扩增 快速检测 结核分枝杆菌复合群 卡介苗 

分 类 号：R735[医药卫生—肿瘤]