PCR-SSCP银染法检测结核分枝杆菌rpsL基因突变

摘要

Objective:To study the value of detection of rpsl gene mutation in streptomy cin(SM)-resistance in clinical isolates of mycobacterium tuberculosis by PCR -SSCP,and to expect to set up a r apid detective method of rpsl gene mutation in mycobacterium tuberculosis,Method Eighty seven specimens isolated from patients were detected by PCR-SSCP silver staining,and using strain H 37Rv as a control,Results In 87 mycobacterium tuberculosis clinical isolates,the r psl DNA fragments from 16 drug-susceptible isolates had no mutation in rpsL gene ,48 of 62 sputum specimens showed rpsl gene mutation by PCR-SSCP,positive rate w as 77.4%.Conclusions The rpsL gene mutation is an important molecular mechanism of M.tuberc ulosis resistance to SM.PCR-SSCP might become a rapid detecting method of SM-r esistance of M.tuberculosis.%目的　探讨结核分枝杆菌对链霉素(SM)的耐药分子机制，建立 快速、准确的SM耐药性 检测方法。方法　应用PCR-SSCP银染分析技术，检测87株结核分枝杆菌 临床分离株链霉素耐药 性及rpsL基因变异情况，以结核分枝杆菌H37Rv标准株做对照。结果　25株对SM敏感的临床 分离株rpsL基因未见异常，62株耐SM的临床分离株中48株rpsL基因SSCP分析结果有异常。 结论　rpsL基因突变是SM耐药性的重要分子机制，PCR-SSCP技术可以快速、准确地 检测结核分枝杆菌对SM的耐药性。