多重PCR法检测结核分枝杆菌耐药基因研究

Multiplex polymerase chain reaction for detection of drug resistance genes of mycobacterium tuberculosis

摘要: 目的:建立多重PCR法快速检测结核分枝杆菌耐药基因。方法:采用多重聚合酶链反应(PCR)同时扩增46株结核分枝杆菌临床分离株、48株北京标准菌株、40例耐药及40例非耐药的结核分枝杆菌临床分离菌株的inhA、katG、rpoB、IS1081等基因并采用测序的方法进行验证。结果:所有耐药样本基因(包括利福平、异烟肼耐药基因)及野生型菌株均被成功扩增。结论:应用多重PCR技术可同时快速检测结核分枝杆菌耐药基因。

Abstract: Objective:To establish a rapid detection method of mycobacterium tuberculosis. Methods:Multiplex polymerase chain reaction(PCR) was employed to dectect katG,IS1081,inhA and rpoB genes of mycobacterium tuberculosis. Forty-eight DNA samples were obtained from the national institute for the control of pharmaceutical and biological products (NICPBP), forty drug resistant mycobacterium tuberculosis were isolated and 40 drug sensitive mycobacterium tuberculosis were isolated from the culture-based bacterias and 46 from clinical samples. All results have been confirmed by sequencing. Results:These genes were successfully 1detected in all samples by multiplex PCR. Conclusions:Multiplex PCR can be used to detect target genes of mycobacterium tuberculosis rapidly.