摘要
目的 应用全基因组测序技术分析宁波市利福平耐药结核分枝杆菌(RR-TB)耐药特征,为RR-TB感染患者的临床诊疗及疫情防控提供科学依据。方法 选取宁波市耐药肺结核唯一定点诊疗机构(宁波市第二医院)耐药监测期间保存的2018-2019年59株RR-TB临床分离株,提取总DNA后进行全基因组测序。应用Perl脚本处理原始数据,并通过结核分枝杆菌(MTB)耐药基因数据库确定耐药基因相关突变,同时结合表型药敏试验结果综合分析本地区RR-TB临床分离株对异烟肼(INH)、利福平(RIF)、乙胺丁醇(EMB)、链霉素(SM)、阿米卡星(AM)、卷曲霉素(CAP)、左氧氟沙星(LFX)、丙硫异烟胺(PTO)及对氨基水杨酸(PAS)9种抗结核药物的耐药特征。结果 59株RR-TB菌株INH、RIF的耐药率分别为83.05%、100.00%;基于全基因组测序的59株RR-TB菌株共检测到40种耐药相关基因突变类型,其中以katG基因Ser315Thr(62.71%),rpoB基因Ser450Leu(62.71%),rpsL基因Lys43Arg(50.85%)和embB基因Met306Val(30.51%)为主;59株RR-TB菌株分为2个基因型,分别为L2型(89.83%)和L4型(10.17%);其中,L2型菌株又分为2个亚型,分别为L2.1型菌株(1.89%)和L2.2型菌株(98.11%);L4型菌株又分为3个亚型,分别为L4.2型菌株(16.67%),L4.4型菌株(66.67%)和L4.5型菌株(16.67%);9种抗结核药物的耐药基因主要突变类型在L2型菌株中的占比远高于L4型。结论 宁波地区RR-TB临床分离株对9种抗结核药物的耐药形势较为严峻,主要流行基因型为L2型,需进一步加强耐药结核病的防控工作。
OBJECTIVE To analyze the characterization of drug resistance of rifampicin-resistant Mycobacterium tuberculosis(RR-TB)isolates in Ningbo by whole genome sequencing technology,to provide scientific basis for the clinical diagnosis and treatment of RR-TB patients and epidemic prevention and control.METHODS A total of 59 clinical isolates of RR-TB were isolated from the only designated medical institution for drug-resistant tuberculosis in Ningbo(Ningbo No.2 Hospital)during the period of surveillance of drug resistance between 2018 and 2019.And the whole genomic DNA extracted from clinical isolates were sequenced.The sequencing data were analyzed with an in-house pipeline which can analyze the gene mutations based on MTB drug resistance gene database.The conventional drug susceptibility test was also used to detect the drug-resistant profiles against nine antibiotics,including isoniazid,rifampicin,ethambutol,streptomycin,amikacin,capreomycin,levofloxacin,prothionamide and para-aminosalicylic acid.RESULTS The drug-resistant rates of 59 RR-TB strains to INH and RIF were 83.05%and 100.00%,respectively.A total of 40 drug resistance-associated mutations were detected by whole genome sequencing,and the most common mutation sites were katG Ser315Thr(62.71%),rpoB Ser450Leu(62.71%),rpsL Lys43Arg(50.85%)and embB Met306Val(30.51%).The two genotypes were founded in 59 strains of RR-TB,including lineage 2(L2)(89.83%)and lineage 4(L4)(10.17%).The L2 was subdivided into two subtypes:L2.1(1.89%)and L2.2(98.11%).The L4 was subdivided into three subtypes:L4.2(16.67%),L4.4(66.67%)and L4.5(16.67%).The proportions of the main drug resistance gene mutations against nine anti-tuberculosis drugs in L2 were much higher than those in L4.CONCLUSION The prevalence of RR-TB against nine anti-tuberculosis drugs was serious in Ningbo with the main genotype of L2.It is necessary to strength the prevention and control of RR-TB.
作者
陈琴
车洋
柳正卫
贺天锋
杨天池
桑国鑫
CHEN Qin;CHE Yang;LIU Zheng-wei;HE Tian-feng;YANG Tian-chi;SANG Guo-xin(Ningbo No.2 Hospital,Ningbo,Zhejiang 315010,China;不详)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2023年第16期2458-2462,共5页
Chinese Journal of Nosocomiology
基金
浙江省基础公益研究计划基金资助项目(LTGY23H190001,LTGY23H190002)
浙江省医药卫生科技计划基金资助项目(2022KY1189)。
关键词
耐药
测序
基因组
结核分枝杆菌
基因突变
利福平耐药结核分枝杆菌
Drug resistance
Sequencing
Genome
Mycobacterium tuberculosis
Gene mutation
Rifampicin-resistantMycobacteriumtuberculosis
