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。
【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.
作者
王一霖
郭向辉
彭焜
王乙好
袁羽
刘颂蕊
耿毅
WANG Yilin;GUO Xianghui;PENG Kun;WANG Yihao;YUAN Yu;LIU Songrui;GENG Yi(Institute of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;Sichuan Key Laboratory of Conservation Biology on Endangered Wildlife,Chengdu Research Base of Giant Panda,Chengdu 610081,China)
出处
《四川农业大学学报》
CSCD
北大核心
2024年第3期672-680,共9页
Journal of Sichuan Agricultural University
基金
成都大熊猫繁育研究基金会资助(CPF2017-13、CPF2017-18)。
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
extraintestinal pathogenic E.coli
giant panda
molecular typing
antibiotic resistance
antibiotic resistance mechanism
