福建省耐多药结核分枝杆菌耐药相关基因突变特征

Molecular characteristics of drug-resistance genes related to INH and RFP in Mycobacterium tuberculosis isolated from MDR-TB in Fujian province

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 characteristics of mutations in genes related to rifampin and isoniazid resistance in MDR-TB isolates from clinic.Methods Four gene fragments rpoB,katG,inhA and mabA-inhA of 67 MDR-TB isolates were sequenced and compared with the reference sequences in H37Rv by using PCR sequencing technology from Fuzhou Pulmonary Hospital in Fujian province.Results In 67 MDR-TB isolates,the mutation rate in RRDR of rpoB was 91.04%（61/67）,and mutation rate at the genes related to INH was 79.10%（53/67）.The main SNP sites（88.06%,59/67） are found at 531,526 and 516 loci of the rpoB gene.The SNPs at katG315 and mabA-inhA（-15）are found within 59.70%（40/67）isolates.No mutation was found in any isolates at both inhA and mabA-inhA gene.Conclusion PCR-DNA sequencing technology can be used to detect the sensitivity of MDR-TB isolates to RFP and INH.