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中国防痨杂志 ›› 2000, Vol. 22 ›› Issue (1): 8-11.

• 论著 • 上一篇    下一篇

应用PCR-RFLP分析结核分枝杆菌rpsL基因突变

吴雪琼;张俊仙;庄玉辉;   

  1. 解放军309医院结核病研究室 北京100091;
  • 收稿日期:1997-06-17 修回日期:1997-11-10 出版日期:2000-01-10 发布日期:2000-01-10

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

WU Xueqiong,ZHANG Junxian,ZHUANG Yuhui.   

  1. Tuberculosis Research Laboratory,The 309th Hospital,Beijing 100091.
  • Received:1997-06-17 Revised:1997-11-10 Online:2000-01-10 Published:2000-01-10

摘要: 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?

关键词: 分枝杆菌,结核, 药物耐受性, 聚合酶链反应, 限制性片段长度多态性

Abstract: 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.

Key words: M.tuberculosis, Drug resistance polymerase chain reaction, Restriction fragment length polymorphism