深圳地区结核临床分离菌基因突变与耐药的关系

目的研究深圳地区结核分枝杆菌临床分离株的基因突变与利福平（RFP）、异烟肼（INH）、链霉素（SM）和乙胺丁醇（EMB）耐药之间的关系。方法采用反向斑点杂交技术（RDBHA）对182株结核分枝杆菌临床分离株的rpoB、katG、inhA、rpsL和embB基因突变位点进行检测,并采用L-J比例法检测这些分离株对RFP、INH、SM、EMB的耐药性。结果 182株临床分离株中,总的基因突变率为34.62%（63/182）,其中rpoB基因突变率最高,达24.17%（44/182）。多重耐药结核菌（MDR-TB）占17.03%（31/182）。以L-J比例法作为金标准,采用RDBHA技术检测分别与RFP、INH、SM、EMB耐药相关的rpoB、katG/inhA、rpsL、embB基因突变,其灵敏度分别为88.89%、67.50%、81.82%和64.29%,特异度分别为97.08%、94.37%、97.99%和92.86%。结论深圳地区结核分离株ropB基因突变最普遍,S531L位点是其突变率最高的位点。深圳地区结核分枝杆菌对4个一线抗痨药物耐药现象均较严重,反向斑点杂交技术（RDBHA）可快速检测结核分枝杆菌耐药基因突变,能给临床提供快速用药指导。

Association between gene mutations and the development of drug resistance of Mycobacterium tuberculosis from the clinical isolates from Shenzhen

Objective To investigate possible association between the gene mutations and the development of rifampin （RFP）,isoniazid （INH）,streptomycin （SM）,and ethambutol （EMB） resistance of Mycobacterium tuberculosis isolated from Shenzhen.Methods 182 MTB clinical isolates from Shenzhen were extensively analyzed for their rpoB,katG,inhA,rpsL and embB mutations using the reverse dot blot hybridization assay （RDBHA）.L-J slant proportion method was used to evaluate the susceptibility of the isolates to INH,RFP,SM,and EMB.Results The overall frequency of gene mutation was 34.62% （63/182）,and rpoB had the highest mutation frequency （24.17%,44/182）.The frequency of MDR-TB was 17.03% （31/182） among the 182 isolates.The sensitivity to rpoB,katG/inhA,rpsL,and embB was 88.89%,67.50%,81.82% and 64.29%,respectively;and the specificity was 97.08%,94.37%,97.99% and 92.86%,respectively.Conclusions Mutation of ropB,especially the S531L mutation,is commonly found in the isolates.Most of the isolate were resistant to the four first-line anti-tuberculosis drugs.RDBHA can be used for fast detection of drug resistance.