莫西沙星和氧氟沙星耐药结核分枝杆菌的分子特征研究  

Molecular characterization of Mycobacterium tuberculosis co-resistant to moxifloxacin and ofloxacin

作　　者：肖阳华 吴小翠[2] 杨景卉[2] 郭银娟 余方友 Xiao Yanghua;Wu Xiaocui;Yang Jinghui;Guo Yinjuan;Yu Fangyou(School of Public Health,Nanchang University,Nanchang 330019,China;Department of Clinical Laboratory,Shanghai Pulmonary Hospital,School of Medicine,Tongji University,Shanghai 200433,China)

机构地区：[1]南昌大学公共卫生学院,南昌330019 [2]同济大学附属上海市肺科医院检验科,上海200433

出　　处：《中华微生物学和免疫学杂志》2024年第4期346-352,共7页Chinese Journal of Microbiology and Immunology

基　　金：上海市加强公共卫生体系建设三年行动计划(GWVI-11.2-YQ52)。

摘　　要：目的利用全基因组测序技术分析上海地区莫西沙星和氧氟沙星同时耐药的结核分枝杆菌(Mycobacterium tuberculosis,Mtb)分子特征,为Mtb感染患者的临床诊疗提供科学依据。方法收集同济大学附属上海市肺科医院保存的142株莫西沙星和氧氟沙星同时耐药的Mtb临床分离株。使用Sensititre®MYCOTB plate进行微孔板法药敏试验;Illumina平台对142株Mtb进行测序;TB-Profiler确定其谱系和耐药相关基因的突变位点;Snippy进行细菌核心基因组系统发育分析。结果表型药敏结果显示142株莫西沙星和氧氟沙星耐药的Mtb分离株对异烟肼和利福平的耐药率分别为33.80%(48/142)和22.54%(32/142);76.06%(108/142)的分离株检测到14种氟喹诺酮耐药基因相关突变,其中以gyrA基因Asp94Gly(21.83%,31/142)、Ala90Val(15.49%,22/142)和Asp94Tyr(9.15%,13/142)突变为主。异烟肼、利福平、乙胺丁醇、链霉素、阿米卡星、乙硫异烟胺和卡那霉素的常见耐药突变类型分别为katG Ser513Thr(68.75%,33/48)、rpoB Ser450Leu(53.13%,17/32)、embB Met306Val(44.44%,8/18)、rpsL Lys43Arg(77.50%,31/40)、rrs 1401A→G(100.00%)、fabG1-15C→T(50.00%,4/8)、rrs 1401A→G(100.00%)。142株Mtb分为2个谱系,分别为L2谱系(92.25%,131/142)和L4谱系(7.75%,11/142);其中L2谱系以L2.2.1亚型(80.28%,114/142)为主,L4谱系以L4.5亚型(5.63%,8/142)为主。系统发育分析和单核苷酸多态性分析显示多株L2谱系菌株可能存在流行病学联系。结论本地区莫西沙星和氧氟沙星耐药的Mtb主要流行基因型为L2型,应加强耐药结核病的防控工作。Objective To analyze the molecular characteristics of Mycobacterium tuberculosis(Mtb)strains co-resistant to moxifloxacin and ofloxacin in Shanghai using whole-genome sequencing(WGS),and provide scientific reference for clinical diagnosis and treatment of Mtb infection.Methods This study collected 142 clinical Mtb isolates that were co-resistant to moxifloxacin and ofloxacin at the Shanghai Pulmonary Hospital Affiliated to Tongji University.Sensititre®MYCOTB plates were used for drug susceptibility testing.WGS of the 142 Mtb strains was performed using the Illumina platform.The lineage and mutation sites of drug-resistant genes were determined using TB-Profiler,and bacterial core genome phylogenetic analysis was performed using Snippy.Results Phenotypic drug susceptibility results revealed that among the 142 Mtb isolates,33.80%(48/142)were resistant to isoniazid and 22.54%(32/142)were resistant to rifampicin.Fourteen mutations in fluoroquinolone-resistant genes were identified in 76.06%(108/142)of the isolates,with Asp94Gly(21.83%,31/142),Ala90Val(15.49%,22/142),and Asp94Tyr(9.15%,13/142)mutations in the gyrA gene being the predominant.The common mutations associated with resistance to isoniazid,rifampicin,ethambutol,streptomycin,amikacin,ethionamide,and kanamycin were katG Ser513Thr(68.75%,33/48),rpoB Ser450Leu(53.13%,17/32),embB Met306Val(44.44%,8/18),rpsL Lys43Arg(77.50%,31/40),rrs 1401A→G(100.00%),fabG1-15C→T(50.00%,4/8),and rrs 1401A→G(100.00%),respectively.The 142 Mtb isolates were classified into two lineages,L2 lineage(92.25%,131/142)and L4 lineage(7.75%,11/142),with L2.2.1(80.28%,114/142)and L4.5(5.63%,8/142)being the predominant subtypes of the corresponding lineages.Phylogenetic analysis and single nucleotide polymorphism detection indicated potential epidemiological connections among multiple L2 lineage strains.Conclusions The prevalent Mtb isolates co-resistant to moxifloxacin and ofloxacin in this region belong to L2 lineage.The prevention and control of drug-resistant Mtb should be strengthe

关 键 词：结核分枝杆菌 莫西沙星 氧氟沙星 全基因组测序 耐药基因 

分 类 号：R52[医药卫生—内科学]