上海市污水处理厂携带mcr-1耐药基因肠道杆菌菌株的分离及拷贝数定量分析
Isolation of Enterobacteriaceae strains carrying mcr-1 resistance gene from Shanghai wastewater treatment plants and quantification of their copy number
丰俊 1刘明香 2庄源 1潘淼 1刘倩 1陈涌 1罗嘉远 1费加艺 1吴艺童 1朱彦祺 1张静 2陈敏1
作者信息
- 1. 上海市疾病预防控制中心病原生物检定所,上海 200336
- 2. 上海市虹口区疾病预防控制中心,上海 200082
- 折叠
摘要
[目的]了解上海市污水处理厂进水中携带多黏菌素类耐药基因-1(mcr-1)的情况及肠道杆菌菌株种类,为污水中含有该类耐药基因菌株的分子监测提供技术支持.[方法]对2023年4-6月收集的上海市污水处理厂的进水样品,采用血琼脂和SS培养基分离培养携带mcr-1耐药基因的肠道杆菌菌株.采用质谱分析仪进行菌株鉴定,利用实时荧光定量聚合酶链反应(PCR)检测其mcr-1基因和拷贝数,应用微量肉汤稀释法进行药敏试验.利用SPSS 25.0软件针对不同来源分离得到的携带mcr-1基因的大肠埃希菌(Escherichia coli)的拷贝数进行统计学分析.[结果]从49份污水进水样品中共分离得到14株携带mcr-1耐药基因的肠道杆菌菌株,阳性分离率为28.6%,包括12株非致泻性大肠埃希菌和2株肺炎克雷伯菌(Klebsiella pneumoniae).药敏试验结果显示,14株mcr-1阳性菌株均为多重耐药(MDR)菌.其中所有mcr-1阳性菌株均对亚胺培南和替加环素敏感,对氨苄西林和头孢唑林的耐药率为100%.人源携带的致泻性大肠埃希菌和污水源分离得到的非致泻性大肠埃希菌之间拷贝数的差异无统计学意义(t=0.647,P>0.05).[结论]首次在上海市污水处理厂的进水中分离得到携带mcr-1耐药基因的非致泻性大肠埃希菌和肺炎克雷伯菌,并发现拷贝数与人源携带mcr-1的致泻性大肠埃希菌差异无统计学意义.污水来源分离得到的菌株MDR现象较为严重.应重视和加强mcr-1耐药基因的监测,以有效防控多黏菌素耐药菌的传播和扩散.
Abstract
[Objective]To provide technical support for the molecular surveillance of pathogenic bacteria strains carrying mobile colistin resistance-1(mcr-1)gene isolate from inlet of wastewater treatment plants(WWTP).[Methods]The Enterobacteriaceae strains carrying mcr-1 resistance gene isolate from inlet of WWTP during April 1 to June 30,2023 in Shanghai were cultured on blood-rich and SS culture medium and were identified using a mass spectrometry analyzer.The mcr-1 gene and copy number were detected by real-time fluorescence quantitative PCR.Drug susceptibility test was performed by microbroth dilution method.The copy numbers of Escherichia coli carrying mcr-1 gene isolated from wastewater and human fecel were statistically analyzed by SPSS 25.0.[Results]A total of 14 strains carrying the mcr-1 gene were isolated from 49 WWTP samples,and the positive isolation rate was 28.6%,including 12 non-diarrheal E.coli strains and 2 Klebsiella pneumoniae strains.The drug susceptibility results showed that all 14 strains were multi-drug resistant bacteria.They were all sensitive to imipenem and tigecycline,but were ampicillin-and cefazolin-resistant.There was no significant difference in the copy number between human-sourced diarrheal E.coli and wastewater-sourced non-diarrheal E.coli(t=0.647,P>0.05).[Conclusion]The isolation and identification of strains carrying the mcr-1 gene from inlet of WWTP samples were firstly established in Shanghai.The multi-drug resistance among the isolated strains is severe.To effectively prevent and control the spread of colistin-resistant bacteria,more attention should be paid to the surveillance of mcr-1 gene.
关键词
污水处理厂/mcr-1基因/分离/拷贝数/耐药性/大肠埃希菌Key words
wastewater treatment plant/mcr-1 gene/isolation/copy number/drug resistance/Escherichia coli引用本文复制引用
基金项目
上海市加强公共卫生体系建设三年行动计划(2023-2025)人才项目和重点学科项目(GWVI-11.2-XD28)
上海市加强公共卫生体系建设三年行动计划(2023-2025)人才项目和重点学科项目(GWVI-11.1-09)
上海市虹口区疾病预防控制中心人才培养项目(ZXRC-2022-03)
出版年
2024
国家科技期刊平台
NSTL
万方数据