中文摘要：

为建立一种快速、高敏感、高特异、鉴别结核分枝杆菌和牛分枝杆菌的方法,本研究根据大多数致病性结核分枝杆菌共有序列esat-6,结核分枝杆菌和牛分枝杆菌gyrB共有特异性序列,以及结核分枝杆菌种特异序列mtp40设计6对特异性引物对痰液中的结核分枝杆菌进行检测,并与鉴别培养基分离培养结果以及常规PCR鉴定结果进行比较。实验结果表明,建立的LAMP检测方法具有很高的特异性。可区分致病性结核分枝杆菌和非致病结核分枝杆菌,也可鉴别结核分枝杆菌和牛分枝杆菌。LAMP检测技术的灵敏度比经典PCR技术高100倍左右,可检测到7拷贝/反应。另外,用3种方法同时检测样品发现,LAMP与细菌培养的符合率为90.91%,LAMP与常规PCR检测结果的符合率为100%。LAMP检测方法可以在流行病学调查及现场快速诊断方面广泛应用,并为临床治疗提供依据。

英文摘要：

To develop an efficient,sensitive and specific method for detection of Mycobacterium tuberculosis and Mycobacterium bovis,specifc primers were designed for a multi-site loop-mediated isothermal amplification（LAMP） method by targeting the esat-6 gene,gyrB gene and mtp40 gene. The results of LAMP detection were compared with that of bacterium isolation and PCR detection. The LAMP is highly efficient and specific in detection of M. tuberculosis and M. bovis and could detect 7 copies/reaction,which was 100 times more sensitive than PCR method. The LAMP assay had 90.91 % consistency with bacterium isolation and 100 % consistency with PCR method. This new method could be applicable in clinical laboratory and field surveillance.