大熊猫源肠外致病性大肠杆菌的分子分型、耐药表型与耐药基因分析
Molecular Typing,Antibiotic Resistance Phenotype and Antibiotic Resistance Gene Analysis of Extraintestinal Pathogenic Escherichia coli from Giant Panda Origin
王一霖 1郭向辉 1彭焜 1王乙好 1袁羽 1刘颂蕊 2耿毅1
作者信息
- 1. 四川农业大学动物医学院,成都 611130
- 2. 成都大熊猫繁育研究基地/四川省濒危野生动物保护生物学重点实验室,成都 610081
- 折叠
摘要
[目的]明确四川大熊猫源肠外致病性大肠杆菌(Ex PEC)的耐药性与耐药基因流行情况,为大熊猫源Ex PEC感染的临床用药提供参考.[方法]从四川省绵阳市和雅安市野外死亡大熊猫的肠外器官采集病料,进行病原菌的分离,通过16S rRNA基因序列分析与Ex PEC多重PCR鉴定,继而对分离的Ex PEC进行系统进化群分群、MLST分型和PFGE分型,并采用K-B法与Wafergen Smartchip超高通量qPCR进行耐药表型测定和耐药基因分析.[结果]分离出9株Ex PEC,系统进化群分别为B2群和A群;MLST和PFGE分型均为3种分子型,其中MLST为ST555、U10和ST127型,PFGE为Ⅰ、Ⅱ和Ⅲ型;9株Ex PEC对多黏菌素B、亚胺培南、左氧氟沙星与萘啶酸敏感率达100%;对β-内酰胺类、氨基糖苷类、四环素类等9类抗生素耐药基因(ARGs)的检测发现,β-内酰胺类和多重耐药类ARGs检出率较高.[结论]分离鉴定的9株大熊猫源Ex PEC系统进化群主要为B2群,MLST和PFGE分型主要为ST127/PFGE基因Ⅰ型,具有明显的多重耐药性,且β-内酰胺类和多重耐药类ARGs检出率较高,主要通过抗生素失活(antibiotic deactivate)和抗生素外排泵(efflux pump)两种耐药机制介导耐药性.
Abstract
[Objective]This study aimed to clarify the antibiotic resistance and the prevalence of antibi-otic resistance genes of extraintestinal pathogenic Escherichia coli(Ex PEC)of giant panda origin in Sich-uan province,and to provide a reference for the clinical antibiotic use to the giant panda infected by Ex PEC.[Method]The pathogenic bacteria were isolated from the extraintestinal organs of wild dead giant pandas in Mianyang and Yaan,Sichuan Province.16S rRNA gene sequence analysis and Ex PEC multi-plex PCR were used for identification,and then the phylogenetic groups,MLST typing and PFGE typing were performed for the isolated Ex PEC.Kirby-Bauer method and Wafergen Smartchip ultra-high throughput qPCR were used to detect antibiotic resistance phenotypes and analyze antibiotic resistance genes.[Result]9 strains of Ex PEC were isolated,and the phylogenetic groups were respectively B2 and A.MLST and PFGE were divided into 3 molecular types,of which the MLST was of ST555,U10 and ST127 types,and the PFGE was of types Ⅰ,Ⅱ and Ⅲ.9 Ex PEC strains were 100%sensitive to poly-myxin B,imipenem,levofloxacin and nalidixic acid.The detection of 9 types of antibiotic resistance genes(ARGs)such as β-lactam,aminoglycosides and tetracyclines resistance presented that the detec-tion rates of β-lactam and multi-drug resistance ARGs were higher.[Conclusion]The phylogenetic group of the 9 strains isolated and identified was mainly B2 group,and the MLST and PFGE types were mainly ST127(MLST)and I(PFGE).The isolated strains exhibited multidrug resistance,with high de-tection rates of β-lactams and multidrug-resistant ARGs.Antibiotic resistance mechanism is mainly me-diated by antibiotic deactivate and efflux pump.
关键词
肠外致病性大肠杆菌/大熊猫/分子分型/耐药性/耐药机制Key words
extraintestinal pathogenic E.coli/giant panda/molecular typing/antibiotic resistance/antibiotic resistance mechanism引用本文复制引用
基金项目
成都大熊猫繁育研究基金会项目(CPF2017-13)
成都大熊猫繁育研究基金会项目(CPF2017-18)
出版年
2024
国家科技期刊平台
NSTL
万方数据