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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。
Objective To investigate the relationship between gene mutations in clinical isolates of Mycobacterium tuberculosis in Shenzhen and rifampicin (RFP), isoniazid (INH), streptomycin (SM) and ethambutol (EMB). Methods The mutation sites of rpoB, katG, inhA, rpsL and embB in 182 clinical isolates of Mycobacterium tuberculosis were detected by reverse dot blot hybridization (RDBHA). The LJ ratio was used to detect the effect of RFP, INH , SM, EMB resistance. Results Among 182 clinical isolates, the total gene mutation rate was 34.62% (63/182). The highest mutation rate of rpoB gene was 24.17% (44/182). MDR-TB accounted for 17.03% (31/182). Using LJ proportional method as the gold standard, RDBHA was used to detect the mutations of rpoB, katG / inhA, rpsL and embB related to resistance to RFP, INH, SM and EMB respectively, with the sensitivity of 88.89%, 67.50% and 81.82% 64.29% and specificity of 97.08%, 94.37%, 97.99% and 92.86% respectively. Conclusion The mutations of ropB gene in tuberculosis isolates in Shenzhen are the most common. The S531L locus is the site with the highest mutation frequency. Mycobacterium tuberculosis in Shenzhen in four first-line anti-tuberculosis drug resistance are more serious, reverse dot blot hybridization (RDBHA) can detect mutations in Mycobacterium tuberculosis resistance gene, can provide rapid clinical guidance.