耐多药结核病耐药分子机制的研究  

Study on the molecular mechanism of multi-drug resistance in clinical isolates of Mycobacterium tuberculosis

作　　者：吴雪琼 庄玉辉 张俊仙 张晓刚 何秀云 李国利 

机构地区：[1]解放军第三O九医院结核病研究室

出　　处：《中华结核和呼吸杂志》1997年第6期332-335,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的研究耐多药结核分支杆菌耐药的分子机理，建立快速检测耐药基因型的分子药敏试验方法。方法通过ＰＣＲ和ＰＣＲ－ＳＳＣＰ分析结核分支杆菌耐多药临床分离株的ｒｐｏＢ、ｒｐｓＬ、ｋａｔＧ基因和ｉｎｈＡ调节序列。结果ＰＣＲ分析耐药基因的敏感性为１～１０ｐｇＤＮＡ；除ｒｐｏＢ基因引物ＰＣＲ扩增为属特异性外，余耐药基因引物ＰＣＲ扩增都具有较高的特异性。２０株耐多药分离株中，９０％有２种以上耐药遗传标志改变，１０％只有ｒｐｏＢ基因突变。结论耐多药是各种药物靶基因逐步突变所致，少数由一个耐多药位点突变所致，通过ＰＣＲ、ＰＣＲ－ＳＳＣＰ可简便、快速地检测大部分结核分支杆菌耐多药分离株的耐药基因型。Objective To study the molecular mechanism of multi-drug resistance in M. Tuberculosis, and to develop a new method for detecting genes related with multi-drug resistance. Method The ropB, rpsL, katG genes and inhA regulatory sequence in clinical isolates of M. tuberculosis were analyzed with PCR and PCRSSCP techniques. Result The sensitivity of amplifing the drugresistant genes with PCR was 1～10 pg DNA. Of the 20 multiple resistant strains with reduced sensitivity to streptomycin, rifampin and isoniazid, 90% showed mutations in more than two genetic markers associated with resistance to each of these three drugs, 10% revealed only mutations in rpoB gene. Conclusion Multi-drug resistance in M. tuberculosis could be caused by an accumulation of mutations in chromosomal genes encoding drug targets or an alteration at a single multiple resistance locus. PCR and PCRSSCP techniques might become simple, rapid and reliable diagnostic tests for multi-drug resistance.

关 键 词：结核病 耐多药结核病 耐药分子机制 PCR 

分 类 号：R52[医药卫生—内科学]