医院未处理污水中碳青霉烯类抗生素耐药细菌的流行情况和临床相关性分析  

Prevalence and clinical relevance of carbapenem-resistant bacteria in untreated hospital wastewater

作　　者：钱费楠 翟雅轩 林佳瑶 王靓 朱志宸 闻奕丞 吕晶南 杜鸿[1] Qian Feinan;Zhai Yaxuan;Lin Jiayao;Wang Liang;Zhu Zhichen;Wen Yicheng;Lyu Jingnan;Du Hong(Department of Clinical Laboratory,The Second Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第二医院医学检验中心,苏州1215000

出　　处：《中华预防医学杂志》2024年第4期532-538,共7页Chinese Journal of Preventive Medicine

基　　金：江苏省科技厅重点研发项目(BE2023704)。

摘　　要：探讨医院未处理污水中碳青霉烯类耐药菌的流行情况、基因组特征及临床相关性,为院内评估公共卫生情况、预防交叉感染提供参考依据。收集2023年3月苏州大学附属第二医院污水处理站内未处理污水和26个病区洗手池U型排污管内污水,离心稀释后涂布于含有美罗培南(2 μg/ml)的LB固体平板分离耐药细菌,进行菌种鉴定、药敏分析、碳青霉烯酶基因PCR检测和全基因组测序;对基因组序列进行耐药基因识别,并采用回顾性研究方法,结合多位点序列分型(multilocus sequence typing,MLST)和单核苷酸多态性(single nucleotide polymorphism,SNP)分析,比较其与同季度临床分离株的同源性。结果显示,从医院污水中分离出56株耐碳青霉烯类革兰阴性菌,来源于13个属,其中17株分离自医院总污水,气单胞菌属为最优势属(35.3%,6/17);39株分离自17个病区的污水,假单胞菌属为最优势属(30.8%,12/39)。本院常见污水分离株均为多重耐药细菌,对部分二、三代头孢菌素类药物耐药率可达100%。来源于污水分离株的碳青霉烯酶基因共8种,包括 blaKPC、 blaNDM、 blaIMP、 blaVIM、 blaIND、 blaOXA-58-like、 blaOXA-48-like和 blaOXA-427-like。共39株污水分离株携带碳青霉烯酶基因,医院总污水中携带 blaKPC-2细菌的分离率(35.3%,6/17)最高,病区污水中携带 blaIMP-8细菌的分离率(31.8%,7/22)最高。14株污水分离株同时携带两种碳青霉烯酶基因,共6种组合型。本院还发现了 blaIMP-101新亚型。筛选4株污水分离株和11株临床分离株纳入SNP分析,其中临床和污水来源的2株ST11型肺炎克雷伯菌仅有15个SNP差异,具有高度同源性。综上,医院未处理污水中存在多种多重耐药的条件致病菌,具有在环境中散播耐药基因的潜在风险;医院污水和临床分离的高度同源肺炎克雷伯菌表明医院污水与临床感染的紧密联系。医院需加强对污水环境中耐药细菌和耐药基�To analyze the prevalence,genomic characteristics and clinical relevance of carbapenem-resistant bacteria in untreated hospital wastewater,and to provide a reference basis for in-hospital assessment of public health situation and prevention of cross-infection.In March 2023,untreated wastewater in the wastewater treatment station of the Second Affiliated Hospital of Soochow University and wastewater in the U-shaped wastewater pipes of the hand-washing sinks in 26 wards were collected,centrifuged and diluted,and the drug-resistant bacteria were isolated by using LB solid plates containing meropenem(2μg/ml)for species identification,drug sensitivity analysis,carbapenenase gene PCR detection and whole genome sequencing.The genome sequence was identified for drug resistance genes.Retrospective research was used,combining multilocus sequence typing(MLST)and single nucleotide polymorphism(SNP)analysis,to compare their homology with clinical isolates of the same quarter.The results showed that 56 carbapenem-resistant gram-negative bacteria were isolated from hospital wastewater,originating from 13 genera,of which 17 were isolated from the total hospital wastewater,with Aeromonas spp.as the most dominant genus(35.3%,6/17),and 39 were isolated from the wastewater of 17 wards,with Pseudomonas spp.as the most dominant genus(30.8%,12/39).All common wastewater isolates from our hospital were multidrug-resistant bacteria,with up to 100%resistant to some second-and third-generation cephalosporins.A total of 8 carbapenemase genes originated from wastewater isolates,including bla KPC,bla NDM,bla IMP,bla VIM,bla IND,bla OXA-58-like,bla OXA-48-like,and bla OXA-427-like.39 wastewater isolates carried the carbapenemase genes,and the total wastewater of the hospital carried the highest isolation rate of bla KPC-2 bacteria(35.3%,6/17)and the highest isolation rate of bla IMP-8 bacteria(31.8%,7/22)were found in the wastewater from 26 wards.14 wastewater isolates were found to carry both carbapenemase genes,with a total of 6 combinations

关 键 词：污水 碳青霉烯酶 细菌耐药 

分 类 号：R446.5[医药卫生—诊断学]