浙江省耐药结核分枝杆菌katG、rpoB、embB基因突变特点研究

Study on The Characteristics of Mutations on Mycobacterium Tuberculosis katG、rpoB、embB Gene in Zhejiang Province

目的了解浙江省耐异烟肼、利福平、乙胺丁醇结核分枝杆菌katG、rpoB、embB基因突变特点。方法以结核分枝杆菌标准株H37RV为对照,应用聚合酶链反应——单链构象多态性(PCR-SSCP)技术、聚合酶链反应——直接测序(PCR-DS)技术检测100株耐药结核分枝杆菌临床分离株katG、rpoB、embB基因。结果经PCR-DS检测katG基因突变率达54.8%,以Ser315Thr突变最常见(40.3%);在耐利福平菌株中,rpoB基因突变率达88.9%,以526、531位氨基酸改变最常见,总突变率达77.8%(63/81);在耐乙胺丁醇菌株中,embB基因突变率达60%,以Met306VaI改变最常见(28.9%)。结论PCR-SSCP技术操作简单、检测快速,可作为临床耐药检测的辅助手段;浙江省结核菌耐药基因突变位点同国内研究基本一致,各位点突变形式所占比例有自身特点。

Objective To elucidate the characteristics of katG, rpoB and embB mutations of mycobacterium tuberculosis in Zhejiang province. Methods Compared with the standard strain H37RV, 100 drug - resistant strains were detected by PCR - SSCP and analyzed by direct sequencing technique. Results The mutation rate of katG is 54.8% and the most familiar site is 315 （Ser→Thr） ;88.9% rifampin resistant strains were detect to have mutations, which mainly located at 526 site and 531 site; in the ethambutol resistant strains, the mutation rate is 60% , in which the form of Met306VaI is 28.9%. Conclusions PCR - SSCP technology is a rapid and easy method to detect gene mutations and it could be used to estimate the drug resistance accessorily. The mutation sites of drug resistance strains correspond with current studies conducted in our country, but the proportion of each mutation form has our own characteristics in the study.