应用PCR-RFLP分析结核分枝杆菌rpsL基因突变

Analyzing the mutations of rpsL gene in Mycobacterium tuberculosis clinical isolates by PCR-RFLP

目的 了解结核分枝杆菌链霉素(SM)耐药基因突变情况,建立快速检测耐药突变株的方法。方法 通过PCR-RFLP分析62株结核分枝杆菌临床分离株的rpsL基因突变的部位和性质。结果 以H37RV标准株为对照,13株敏感株的rpsL基因有MboⅡ酶切位点;37株耐SM分离株中,31株(838%)无MboⅡ酶切位点;12株耐其它抗结核药物株中,也有2株无MboⅡ酶切位点。结论 大多数结核分枝杆菌SM耐药分离株的核糖体S12蛋白编码基因(rpsL)第43位密码子有点突变。PCR-RFLP可能成为快速检测结核分枝杆菌耐药突变株的一种新方法。

Objective To observe the mutations of rpsL gene in M.tuberculosis streptomycin-resistant isolates,and to develop a new method for detecting drug resistance.Method Analyzing the mutations of rpsL gene codon 43 in M.tuberculosis clinical isolates with PCR-RFLP.Results M.tuberculosis strain H 37 R V was used as a control.The rpsL gene fragments from 13 drug-sensitive strains could be digested by restriction endonuclease MboⅡ.31(83.8%)of 37 streptomycin resistant strains were not restricted by MboⅡ.2 of 12 non-streptomycin-resistant strains hadn't also sites of MboⅡ.Conclusions Most M.tuberculosis streptomycin-resistant strains could be observed the mutations situated at codon 43 of genes encoding the ribosomal S12 protein (rpsL).PCR-RFLP might become a simple,rapid and reliable means of detecting drug resistance in clinical isolates.