为探讨新疆地区结核分枝杆菌对链霉素(streptomycin,STR)产生耐药性的分子机制,建立直接快速检测结核杆菌STR药物敏感性的方法。应用聚合酶链反应-单链构象多态性(PCR-SSCP)银染分析技术,检测62株结核杆菌临床分离株链霉素耐药性及rpsL和rrS基因变异情况,以结核分枝杆菌标准株(H37Rv)为对照。结果发现62株结核杆菌临床分离株中,32株STR敏感株的rpsL和rrS基因未见SSCP泳动异常,30株STR耐药株中,23株(76.7%)中rpsL(21株)和rrS(5株)基因出现SSCP泳动异常,并且3株出现了rpsL和rrS双基因的SSCP泳动异常。因此,rp-sL和rrS基因突变可能是结核分枝杆菌对链霉素产生耐药性的重要分子机制,PCR-SSCP银染技术可作为快速检测结核杆菌STR药物敏感性的方法。

To study the molecular mechanism of M.tuberculosis resistance to streptomycin,and to set up a method for direct,rapid detection of streptomycin-resistance mutations in M.tuberculosis.Method The rpsL and rrS genes of 62 M.tuberculosis clinical isolates were detected by PCR-SSCP,Strain H37Rv was used as a control.Results In 62 M.tuberculosis clinical isolates,the rpsL and rrS PCR fragments form 32 drug-susceptible isolates had no differences in rpsL and rrS PCR profiles with strain H37Rv.In 30 streptomycin-resistant isolates,23 mol/L.tuberculosis clinical isolates(76.7%)showed apparent differences in SSCP profiles,including 21 mol/L.tuberculosis clinical isolates of rpsL gene mutation,5 mol/L.tuberculosis clinical isolates of rrS gene mutation and 3 mol/L.tuberculosis clinical isolates of rpsL and rrS gene mutation simultaneously.Conclusions The rpsL and rrS gene mutation maybe an important mechanism of M.tuberculosis resistance to streptomycin.PCR-SSCP might be used as a simple,rapid and reliable diagnostic method for M.tuberculosis drug resistance.