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目的初步分析福建耐多药结核病(multi-resistant tuberculosis,MDR-TB)临床分离结核分枝杆菌利福平和异烟肼的耐药相关基因突变情况。方法自福建福州肺科医院收集耐多药结核分枝杆菌临床分离菌株,采用PCR测序技术测定利福平和异烟肼耐药相关基因,包括rpoB、katG、inhA和mabA-inhA,并与标准菌株基因序列比对,分析基因突变特点。结果共对67株MDR-TB菌株进行了分析,91.04%的菌株在rpoB81 bp RRDR区发生突变,其中,88.06%(59/67)的MDR-TB菌株在rpoB531、526和516位发生突变;79.10%的菌株在katG、inhA或mabA-inhA上发生突变,59.70%(40/67)的菌株在katG315或者mabA-inhA(-15)位发生突变,没有检测到同时在inhA结构基因和基因调节区发生突变的菌株。结论 PCR-DNA测序分析技术可以用于检测MDR-TB临床分离菌株对RFP和INH的药物敏感性。
OBJECTIVE: To analyze the mutations of resistance-related genes in rifampicin and isoniazid-resistant Mycobacterium tuberculosis isolates from Fujian multidrug-resistant tuberculosis (MDR-TB). Methods The clinical isolates of multidrug-resistant Mycobacterium tuberculosis were collected from Fuzhou Pulmonary Hospital of Fujian Province. The related genes of rifampicin and isoniazid resistance, including rpoB, katG, inhA and mabA-inhA, were determined by PCR and compared with the standard strains Gene sequence alignment, analysis of gene mutation characteristics. Results A total of 67 strains of MDR-TB were analyzed, 91.04% of them were mutated in rpoB81 bp RRDR, of which 88.06% (59/67) of MDR-TB were mutated at rpoB531, 526 and 516. 79.10 % Of the strains were mutated in katG, inhA or mabA-inhA, 59.70% (40/67) of the strains were mutated in katG315 or mabA-inhA (-15), and no mutation in both inhA structural gene and gene regulatory region A mutated strain. Conclusion The PCR-DNA sequencing technique can be used to detect the drug susceptibility of MDR-TB clinical isolates to RFP and INH.