摘要
目的分析福建省产超广谱β内酰胺酶(ESBLs)宋内志贺菌基因组特征,并对共存于质粒的bla_(CTX-M)耐药基因进行快速鉴定。方法收集福建省6个地区2007—2024年产ESBLs宋内志贺菌26株,微量肉汤法检测抗菌素最小抑菌浓度(MIC);利用二代和三代测序技术进行全基因组测序(WGS),预测耐药基因、质粒、插入序列(IS)和bla_(CTX-M)基因环境,Roary构建聚类树,cgSNP(core genome single nucleotide polymorphism)分析菌株的全球进化地位。S1核酸内切酶脉冲场凝胶电泳(S1-PFGE)、微滴式数字PCR(ddPCR)及质粒接合转移试验确认bla_(CTX-M)在质粒上的定位和水平传播。结果26株产ESBLs宋内志贺菌均为多重耐药菌,全部对头孢噻肟耐药,有6株对阿奇霉素耐药。WGS分析显示,26株菌共携带17种耐药基因、9种质粒复制子、15种插入序列和1种转座子Tn7。bla_(CTX-M-14)、bla_(CTX-M-15)和bla_(CTX-M-64)、ermB和mphA的阳性率分别为61.54%、30.77%、7.69%、11.54%和11.54%,bla_(CTX-M)基因环境可分为4种类型。Roary分析可将26株菌分为4个簇;cgSNP显示,它们均归属于独特的基因型3.7.6中国分支。除1株bla_(CTX-M-14)位于染色体外,所有bla_(CTX-M)分布于3种类型的质粒上,S1-PFGE和ddPCR鉴定结果与测序结果一致。结论福建产ESBLs宋内志贺菌基因组携带大量耐药因子,S1-PFGE和ddPCR可成为检测质粒携带耐药基因的有效工具。
This study analyzed the genomic characteristics of extended spectrum beta lactamase(ESBL)-producing Shigella sonnei(S.sonnei)in Fujian Province,and rapidly located and identified bla_(CTX-M)resistance genes coexisting in the plasmid.Twenty-six isolates of ESBL-producing S.sonnei were collected from Fujian Province during 2007—2024,and the minimum inhibitory concentrations(MICs)of antibiotics were detected with the broth microdilution method.Whole genome sequencing(WGS)was used to predict antibiotic resistance genes,plasmids,insertion sequences,and the bla_(CTX-M)gene environment.A clustering tree was constructed on the basis of Roary analysis,and global evolutionary status of the strains was analyzed according to core genome single nucleotide polymorphisms(cgSNPs).The localization and horizontal transfer of bla_(CTX-M)on the plasmids were confirmed with S1-pulsed field gel electrophoresis(S1-PFGE),microdrop digital PCR(ddPCR),and plasmid conjugation transfer tests.All isolates were ftb multi-drug resistant and resistant to cefotaxime,and six isolates were resistant to azithromycin.WGS analysis identified 17 antibiotic resistance genes,9 types of plasmid replicons,15 types of insertion sequences,and 1 Tn7 transposon among the 26 isolates.The positivity rates of bla_(CTX-M-14),bla_(CTX-M-15),bla_(CTX-M-64),ermB,and mphA were 61.54%,30.77%,7.69%,11.54%,and 11.54%,respectively.Four of bla_(CTX-M)gene environment types were identified.The 26 isolates were divided into four clades through Roary analysis,and cgSNP analysis indicated that these strains all belonged to the unique Chinese clade of genotype 3.7.6.Except for one strain of bla_(CTX-M-14)located on the chromosome,all other bla_(CTX-M)genes were located on three types of plasmid replicons.The S1-PFGE and ddPCR identification results were consistent with the sequencing results.In summary,the genomes of ESBL-producing S.sonnei carry a variety of antibiotic resistance factors,and S1-PFGE combined with ddPCR may be an effective tool for detecting plasmid-carried resistance genes.
作者
康文艳
张小玄
杨劲松
邱玉锋
高亚东
罗朝晨
黄梦颖
KANG Wenyan;ZHANG Xiaoxuan;YANG Jinsong;QIU Yufeng;GAO Yadong;LUO Chaochen;HUANG Mengying(School of Public Health,Fujian Medical University,Fuzhou 350122,China;Fujian Provincial Center for Disease Control and Prevention,Fuzhou 350012,China;Fujian Provincial Key Laboratory of Zoonoses,Fuzhou 350012,China)
出处
《中国人兽共患病学报》
北大核心
2025年第11期1114-1120,共7页
Chinese Journal of Zoonoses
基金
福建省自然科学基金项目(No.2022J01399)
福建省卫生健康科技计划项目(No.2021CXA024)。
关键词
宋内志贺菌
超广谱Β内酰胺酶
全基因组测序
耐药基因
质粒
Shigella sonnei
extended-spectrum beta-lactamases(ESBLs)
whole genome sequencing
antibiotic resistant genes
plasmid
