[用于快速检测结核分枝杆菌的分子信标实时荧光定量PCR技术的研发]
[Development of a molecular beacon real-time PCR for the rapid detection of Mycobacterium tuberculosis].
作者信息
Xu Ya-jun, Zhou Zi-ren, Lin Lin, Liu Qu, Liu Heng-chuan
机构信息
Department of Medical Technology, West China School of Public Health, West China Medical Center, Sichuan University, Chengdu 610041, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Jul;39(4):661-3.
OBJECTIVE
To develop a molecular beacon real-time PCR for rapid detection of Mycobacterium tuberculosis.
METHOD
One set of primers was selected from the IS6110 gene in GenBank and the corresponding molecular beacon probe was designed. The specificity and sensitivity of the developed method were evaluated by tested with 10 different bacteria species. The developed assay were also applied to the diagnosis of tuberculosis.
RESULTS
Only Mycobacterium tuberculosis strains possessing IS6110 gene generated fluorescent signals, and no cross reaction was observed with other 9 bacteria. The detection limit was 4 copies/PCR reaction. 100 Mycobacterium tuberculosis strains were positive tested by Real-time PCR.
CONCLUSION
The established molecular beacon real-time PCR is a rapid, specific and sensitive method, and is a beneficial supplement of traditional methods for the tuberculosis diagnosis.
目的
开发一种用于快速检测结核分枝杆菌的分子信标实时荧光定量聚合酶链反应（PCR）方法。
方法
从GenBank中的IS6110基因中选择一组引物，并设计相应的分子信标探针。通过对10种不同细菌进行检测，评估所开发方法的特异性和灵敏度。所开发的检测方法也应用于结核病的诊断。
结果
只有携带IS6110基因的结核分枝杆菌菌株产生荧光信号，与其他9种细菌未观察到交叉反应。检测限为4拷贝/PCR反应。100株结核分枝杆菌菌株经实时荧光定量PCR检测为阳性。
结论
所建立的分子信标实时荧光定量PCR是一种快速、特异、灵敏的方法，是结核病诊断传统方法有益的补充。
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