浙东地区耐多药结核分枝杆菌耐药特性及分子机制研究  

Study on drug resistance characteristics and molecular mechanism of MDR-TB in eastern Zhejiang Province

作　　者：石庆新[1] 陆如岳[1] 於青峰[1] 涂茜 李蒿蒿 蔡莺莺 周秋菊 周凯 王冬莲 SHI Qingxin;LU Ruyue;YU Qingfeng;TU Xi;LI Haohao;CAI Yingying;ZHOU Qiuju;ZHOU Kai;WANG Donglian(Laboratory Medicine,Taizhou Hospital of Zhejiang Province,Linhai 317000,China)

机构地区：[1]浙江省台州医院检验科,浙江临海317000

出　　处：《中国现代医生》2020年第34期5-8,F0003,共5页China Modern Doctor

基　　金：浙江省医药卫生科技计划项目(2018KY912);浙江省台州市科技计划项目(1802ky20)。

摘　　要：目的对浙东地区耐多药结核分枝杆菌(MDR-TB)耐药性及分子机制进行研究,为治疗耐多药结核病提供理论依据。方法收集2018年1月~2019年12月浙江东部9家结核病定点医院临床分离的结核分枝杆菌。采用比例法检测异烟肼(INH)、利福平(RIF)、氧氟沙星(OFL)、链霉素(SM)、乙胺丁醇(EMB)、阿米卡星(AK)、对氨基水杨酸(PAS)、卷曲霉素(CM)和丙硫异烟胺(TH)对MDR-TB的耐药性。通过基因芯片方法检测耐多药结核分枝杆菌rpoB、katG、inhA突变位点,PCR扩增OFL耐药的MDR-TB的gyr耐药基因并测序。结果耐多药结核分枝杆菌对OFL、SM、EMB、AK、PSA、CM、TH、INH和RIF耐药率分别为38.1%、54.8%、28.6%、11.9%、8.3%、9.5%、13.1%、100.0%和100.0%。耐多药结核分枝杆菌的突变位点为rpoB 511(9例)、rpoB 513(3例)、rpoB 516(3例)、rpoB 526(25例)、rpoB 531(38例)、rpoB 533(2例),KatG 315(71例),inhA-15(4例),KatG 315与inhA-15同时突变(9例)。检测到26株gyrA基因和2株gyrB基因发生突变,突变类型为Thr478Asn、Asn477Thr、Ala90Val、Ser91Pro、Asp94Ala、Asp94His、Asp94Asn和Asp94Gly。结论耐多药结核分枝杆菌利福平耐药以rpoB基因531位点突变为主;异烟肼耐药以KatG基因315位点突变为主;MDR-TB对喹诺酮类药物的耐药机制以gyrA基因Ala90Val、Ser91Pro、Asp94Gly突变类型为主。Objective To study the drug resistance and molecular mechanism of multi-drug resistant mycobacterium tuberculosis(MDR-TB)in eastern Zhejiang province,and to provide theoretical basis for the treatment of MDR-TB.Methods Mycobacteria tuberculosis that were clinically isolated in 9 designated tuberculosis hospitals in eastern Zhejiang province from January 2018 to December 2019 were collected.The drug resistance of MDR-TB to isoniazid(INH),rifampicin(RIF),ofloxacin(OFL),streptomycin(SM),ethambutanol(EMB),amikacin(AK),para-aminosalicylic acid(PAS),capreomycin(CM)and propyl thionisocyanamine(TH)was detected by the ratio method.The rpoB,katG and inhA mutation sites of MDR-TB were detected by the gene chip method.The gyr resistance gene of OFL-resistant MDR-TB was amplified by PCR and sequenced.Results The drug resistance rates of MDR-TB to OFL,SM,EMB,AK,PSA,CM,TH,INH and RIF were 38.1%,54.8%,28.6%,11.9%,8.3%,9.5%,13.1%,100.0%and 100.0%,respectively.The mutational sites of MDR-MTB were rpoB 511(9 cases),rpoB 513(3 cases),rpoB 516(3 cases),rpoB 526(25 cases),rpoB 531(38 cases),rpoB 533(2 cases),KatG 315(71 cases),inhA-15(4 cases),KatG 315 and inhA-15 simultaneously(9 cases).Mutations were detected in 26 gyrA genes and 2 gyrB genes,and the mutation types were Thr478Asn,Asn477Thr,Ala90Val,Ser91Pro,Asp94Ala,Asp94His,Asp94Asn and Asp94Gly.Conclusion The major mutation site of genes in MDR-TB resistant to rifampicin is rpoB 531.The major mutation site of genes in MDR-TB resistant to isoniazid is KatG 315.The resistance mechanism of MDRTB to quinolones is mainly gyrA gene Ala90Val,Ser91Pro and Asp94Gly mutations.

关 键 词：结核分枝杆菌 耐多药 基因芯片 基因突变 

分 类 号：R52[医药卫生—内科学]