高芬, 陈涌, 庄源, 陈洪友, 罗嘉远, 张曦, 陈敏. 上海地区碳青霉烯耐药肠杆菌科细菌的分子生物学特性研究[J]. 上海预防医学, 2021, 33(7): 599-604. DOI: 10.19428/j.cnki.sjpm.2021.20742
引用本文: 高芬, 陈涌, 庄源, 陈洪友, 罗嘉远, 张曦, 陈敏. 上海地区碳青霉烯耐药肠杆菌科细菌的分子生物学特性研究[J]. 上海预防医学, 2021, 33(7): 599-604. DOI: 10.19428/j.cnki.sjpm.2021.20742
GAO Fen, CHEN Yong, ZHUANG Yuan, CHEN Hong-you, LUO Jia-yuan, ZHANG Xi, CHEN Min. Molecular biological characteristics of carbapenem-resistant Enterobacteriaceae in Shanghai[J]. Shanghai Journal of Preventive Medicine, 2021, 33(7): 599-604. DOI: 10.19428/j.cnki.sjpm.2021.20742
Citation: GAO Fen, CHEN Yong, ZHUANG Yuan, CHEN Hong-you, LUO Jia-yuan, ZHANG Xi, CHEN Min. Molecular biological characteristics of carbapenem-resistant Enterobacteriaceae in Shanghai[J]. Shanghai Journal of Preventive Medicine, 2021, 33(7): 599-604. DOI: 10.19428/j.cnki.sjpm.2021.20742

上海地区碳青霉烯耐药肠杆菌科细菌的分子生物学特性研究

Molecular biological characteristics of carbapenem-resistant Enterobacteriaceae in Shanghai

  • 摘要:
    目的分析上海市腹泻患者肠道碳青霉烯耐药肠杆菌科细菌(CRE)的耐药特征和分子分型,为该耐药菌的监测及防控提供依据。
    方法收集上海市腹泻病监测系统23家哨点医院2018年1月—2019年12月肠道门诊患者的粪便或肛拭子样本800份,采用含1 μg/μL美罗培南的麦康凯琼脂培养基分离CRE菌株。使用VITEK-2 Compact全自动细菌鉴定系统和VITEK MS质谱系统进行菌种鉴定。采用微量肉汤稀释法检测菌株的最小抑菌浓度(MIC)。采用多位点序列分型(MLST)及脉冲场凝胶电泳(PFGE)对耐药菌株进行同源性分析。通过接合实验研究耐药基因的可转移性。采用全基因组测序进行耐药分子特征研究。
    结果从800份粪便或肛拭子标本中分离出7株CRE菌株,皆为产新德里金属β-内酰胺酶(NDM)的多重耐药大肠埃希菌,对多种临床常用的抗生素均耐药。分子分型结果显示,CRE菌株序列型别均不相同,PFGE呈现多样性。blaNDM基因均为阳性,包括blaNDM-5blaNDM-13,主要型别为blaNDM-5,并可通过接合转移至大肠埃希菌EC600。
    结论肠道分离碳青霉烯耐药菌株均为多重耐药大肠埃希菌,其耐药机制均为产NDM,同时携带blaNDM及其他多种耐药基因。MLST显示菌株属于不同的克隆型。耐药基因可通过接合转移实现耐药基因的水平转移。

     

    Abstract:
    ObjectiveTo investigate the antimicrobial resistance characteristics of carbapenem-resistant Enterobacteriaceae (CRE) isolated from outpatients with diarrhea in Shanghai, and provide support for surveillance, prevention and control of CRE.
    MethodsA total of 800 fecal swabs of the outpatients with diarrhea were collected from 23 sentinel hospitals for diarrhea pathogen surveillance in Shanghai from January 2018 to December 2019. The drug-resistant strains were isolated using MacConkey plates containing 1 μg/μL meropenem. The collected strains were identified preliminarily by the VITEK-2 Compact system and VITEK mass spectrometry. The minimum inhibitory concentration (MIC) of the strain was determined by the broth microdilution method. The multi-locus sequence typing (MLST) method and pulsed-field gel electrophoresis (PFGE) were used to analyze the homology of drug-resistant strains. The transferability of the resistance gene was investigated by a junction experiment. High-throughput sequencing was used to characterize the isolates.
    ResultsSeven non-repetitive CRE isolates were multi-drug resistant carbapenem-resistant Escherichia coli (CREC) strains that produce New Delhi metallo-β-lactamase (NDM) with resistance to several commonly used antibiotics in clinical therapy. The molecular typing results showed that the CRE strains had different sequence types, and diverse PFGE patterns. The stains were all positive for blaNDM genes, including blaNDM-5 and blaNDM-13, with blaNDM-5 as the main type. The carbapenem-resistant genes could be transferred to EC600 by conjugation.
    ConclusionThe intestinal carbapenem-resistant strains in this study are all NDM-producing Escherichia coli. The isolates carried blaNDM and other resistance genes. The MLST analysis showed that they belonged to different cloning types. Antimicrobial resistance genes could be horizontally transferred to EC600 by conjugation.

     

/

返回文章
返回