耐多药结核分支杆菌突变基因检测

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16S rRNA聚合酶链反应单链的构象多态性（polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)方法对30株耐多药分离株和20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16S rRNA PCR-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，56．7％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，快速地检测大部分耐多药结核病的耐药基因。

GENE MUTATION DETECTION IN MULTIDRUG RESISTANT MYCOBACTERIUM TUBERCULOSIS

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