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 PCRSSCP techniques. Result The sensitivity of amplifing the drugresistant 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 PCRSSCP techniques might become simple, rapid and reliable diagnostic tests for multi-drug resistance.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
1997年第6期332-335,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
关键词
结核病
耐多药结核病
耐药分子机制
PCR
Tuberculosis, multidrugresistant Polymerase chain reaction Polymorphism, single-strand conformational Mycobacterium tuberculosis
