多耐药奇异变形杆菌的耐药基因特征及基因组溯源分析  

Drug resistance gene characteristics and genomic traceability of multidrug resistant Proteus mirabilis

作　　者：张芸 王爽[2] 刘璐 陈玉贞[2] 孙高翔 刘子卿 王安阁 寇增强[2] 王加坤 Zhang Yun;Wang Shuang;Liu Lu;Chen Yuzhen;Sun Gaoxiang;Liu Ziqing;Wang Ange;Kou Zengqiang;Wang Jiakun(School of Public Health and Health Management,Shandong First Medical University(Shandong Academy of Medical Sciences),Jinan 250117,China;Infectious Disease Control Center,Shandong Center for Disease Control and Prevention,Jinan 250014,China;Hospital Infection Management Office,Provincial Hospital Affiliated to Shandong First Medical University(Shandong Academy of Medical Sciences),Jinan 250021,China;Office of Jinan Center for Disease Control and Prevention,Jinan 250021,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)公共卫生与健康管理学院,济南250117 [2]山东省疾病预防控制中心传染病防制所,济南250014 [3]山东第一医科大学(山东省医学科学院)附属省立医院医院感染管理办公室,济南250021 [4]济南市疾病预防控制中心中心办公室,济南250021

出　　处：《中华传染病杂志》2024年第7期416-423,共8页Chinese Journal of Infectious Diseases

基　　金：山东省医药卫生科技项目(202312050986);山东省公共卫生学会科研项目(SGWXH202326、SGWXH202310);山东省青年创新基金项目(QC-202306)。

摘　　要：目的探究山东省泰安市与莱州市门诊腹泻病例来源奇异变形杆菌(PM)的耐药情况及基因组特征。方法收集2021年1月至2022年12月山东省泰安市与莱州市43家哨点医院收治的510例急性腹泻患者的510份粪便样本,通过直接接种培养并分离获得变形杆菌,采用微量肉汤稀释法进行药物敏感试验,通过全基因组测序及生物信息分析获得耐药基因和毒力基因信息,揭示bla OXA-1与bla CTX-M-65基因周围遗传环境。采用RidomSeqSphere+软件通过自定义的方式对本研究菌株与100株从美国国家生物技术信息中心(NCBI)数据库下载的PM分离株进行单核苷酸多态性分析(SNP)与核心基因组多位点序列分型(cgMLST),探究菌株间系统进化关系。结果510份粪便样本共分离获得35株变形杆菌,其中31株PM,检出率为6.08%(31/510),4株普通变形杆菌。PM的多重耐药率为100.00%(31/31)。35株分离株共携带43种耐药基因类型,13株PM携带bla_(OXA-1),6株PM同时携带bla_(OXA-1)与bla_(CTX-M-65)基因,15株PM至少携带15种抗生素耐药基因(ARG)。毒力基因包括ureA、mrpA、ZapA、hpmA等。bla_(OXA-1)与bla_(CTX-M-65)基因周围存在Tn3、ISL3、IS6等移动遗传元件。cgMLST与SNP聚类结果具有一致性,莱州市分离株2022LZ41与泰安市分离株2022TA018、2022TA017、2022TA019聚集在一起,等位基因差异数为0~2;莱州市分离株2022LZ40与CRK0056(人,美国,2015年)参考株具有高度的遗传关系。结论从腹泻患者中检出的PM均为多重耐药菌,且携带大量的耐药基因和毒力基因,可移动遗传元件的存在可能导致耐药基因的水平转移。ObjectiveTo investigate the drug resistance status and genomic characterization of Proteus mirabilis(PM)isolated from outpatient cases with diarrhea in Tai′an City and Laizhou City,Shandong Province.MethodsA total of 510 fecal samples were collected from 510 patients with acute diarrhea admitted to 43 sentinel hospitals in Tai′an City and Laizhou City,Shandong Province,between January 2021 and December 2022.The samples were cultured and isolated to identify Proteus spp.by direct inoculation,the drug susceptibility testing was performed by microbroth dilution method,and resistance genes and virulence genes were obtained by whole genome sequencing and bioinformatic analysis,thereby revealing the genetic environment surrounding the bla OXA-1 and bla CTX-M-65 genes.Single nucleotide polymorphism(SNP)analysis and core genome multilocus sequence typing(cgMLST)were conducted on the current strains and 100 PM isolates downloaded from the National Center for Biotechnology Information(NCBI)database via customizable methods utilizing RidomSeqSphere+software,with the objective of exploring the phylogenetic relationships among the strains.ResultsA total of 35 strains of Proteus were isolated from 510 fecal samples,including 31 strains of PM with a detection rate of 6.08%(31/510)and four strains of Proteus vulgaris.The multidrug resistance rate of PM was 100.00%(31/31).The 35 isolates carried a total of 43 resistance genotypes.Thirteen strains of PM carried bla_(OXA-1),six strains carried both bla_(OXA-1)and bla_(CTX-M-65),and 15 PM strains carried at least 15 antibiotic resistance genes(ARGs).The virulence genes included ureA,mrpA,ZapA,hpmA and so on.bla OXA-1 and bla CTX-M-65 genes were surrounded by mobile elements such as Tn3,ISL3 and IS6.cgMLST showed consistency with the SNP clustering results.Isolate 2022LZ41 from Laizhou City clustered with isolates 2022TA018,2022TA017 and 2022TA019 from Tai′an City,with the number of allelic differences ranging from zero to two,and the Laizhou City isolate 2022LZ40 was highly ge

关 键 词：奇异变形杆菌 全基因组测序 耐药基因 碳青霉烯类耐药 

分 类 号：R516[医药卫生—内科学]