摘要
目的研究耐多药结核分支杆菌耐药分子机制 ,建立快速检测耐药基因型的分子药敏试验方法。方法通过 16SrRNA聚合酶链反应 单链构象多态性 (polymerasechainreaction singlestrandconformationpolymorphism ,PCR SSCP)方法对 30株耐多药分离株和 2 0株药物敏感株进行初步分子菌种鉴定 ;通过PCR SSCP分析 30株耐多药结核病临床分离株的rpoB、katG、rpsL基因。 结果经 16SrRNAPCR SSCP菌种鉴定 ,所分析菌株均为结核分支杆菌 ;30株耐多药分离株经SSCP分析 ,5 6 .7%存在rpoB基因突变 ,43 .3 %有katG基因突变 ,91.3 %有rpsL基因突变。 结论rpoB、rpsL、katG基因突变分别是结核分支杆菌耐利福平、链霉素、异烟肼的分子机制 ,耐多药结核病是药物靶基因突变所致。通过PCR SSCP可简便。
ObjectiveTo research the molecular mechanisms of drug resistance in multidrug resistant mycobacterium(M.) tuberculosis,and to set up a new method to detect the drug resistance of M.tuberculosis.MethodsBy amplifying 16S rRNA gene with PCR SSCP 30 multidrug resistant isolates and 20 sensitive isolates were analyzed.The rpoB,katG,rpsL genes in 30 multidrug resistance M.tuberculosis were analyzed with PCR and PCR SSCP.ResultsAll of isolates were identified as M.tuberculosis by 16S rRNA PCR SSCP analysis. Among 30 multidrug resistant M.tuberculosis , rpoB, katG, rpsL gene mutation rates were 56.7 % , 43.3 % , 91.3 % respectively .ConclusionAlterations in rpoB, rpsL, katG gene may be the important mechanisms of M.tuberculosis resistance to rifampin,streptomycin and isoniazid.PCR SSCP is going to become a simple,rapid diagnostic method for the determination of multidrug resistance M.tuberculosis. MeSH mycobacterium,tuberculosis;tuberculosis,multidrug resistant;polymerase chain reaction;polymorphism,single strand conformatonal
出处
《河北医科大学学报》
CAS
2001年第5期276-279,共4页
Journal of Hebei Medical University
关键词
耐多药结核分支杆菌
突变基因
检测
聚合酶链反应
mycobacterium,tuberculosis
tuberculosis,multidrugresistant
polymerase chain reaction
polymorphism,singlestrand conformatonal
