上海市浦东新区致泻性大肠埃希菌病原谱的建立和分析  

Establishment and analysis for pathogentic spectrum of diarrhoeagenic Escherichia coli in Pudong District of Shanghai

作　　者：赵冰[1] 朱林英[1] 黄红[1] 孙乔[1] 

机构地区：[1]上海市浦东新区疾病预防控制中心微生物科,上海200136

出　　处：《职业与健康》2016年第10期1327-1331,共5页Occupation and Health

基　　金：国家科技重大专项(2012ZX10004201)

摘　　要：目的建立基于分子生物学技术结合传统分离方法检测5种致泻性大肠埃希菌(DEC),并应用于浦东新区腹泻人群的DEC监测,从而构建DEC感染病原谱,以了解浦东新区感染性腹泻患者DEC的感染情况。方法从GenBank基因数据库中筛选5种DEC毒力基因序列,合成引物探针,优化设计TaqMan实时荧光PCR法结合传统分离方法,最终获得单克隆菌株。应用该方法对2014年的11家腹泻监测点1 846份粪便拭子进行DEC检测。结果 1 846件粪便拭子中DEC总阳性率为14.90%(275/1 846),其中检出致病性大肠埃希菌102株(5.53%),产毒性大肠埃希菌102株(5.53%),侵袭性大肠埃希菌3株(0.16%),聚集性大肠埃希菌68株(3.68%)。结论 Taq Man实时荧光聚合酶链反应(PCR)结合传统分离方法分离DEC体现出非常高的准确性和检测效率,同时避免了志贺菌的漏检;为期1年的监测数据显示,DEC已成为细菌性感染性腹泻病原谱中首位病原,其组成结构呈多样性及显著的季节性特点。[Objective]To establish a method for detecting 5 kinds of diarrhoeagenic Escherichia coli（DEC） on the basis of molecular biology technology and traditional separation methods,develop the pathogentic spectrum of DEC by applying the method in DEC monitoring among patients with diarrhea in Pudong District,and know the DEC infection status among patients with diarrhea.[Methods]The virulence gene sequences of 5 kinds of DEC were selected from Gen Bank,to obtain the primers probe. Taq Man real-time fluorescence PCR combined with the traditional separation method was used to acquire the monoclonal strain. 1 846 faecal swabs from 11 diarrhea monitoring points in 2014 were investigated for DECs by the method.[Results]The total positive rate of DEC was 14.90%（275/1 846）among 1 846 faecal swabs. There were 102 strains of pathogenic Escherichia coli（5.53%）,102 strains of enterotoxigenic Escherichia coli（5.53%）,3 strains of enteroinvasive Escherichia coli（0.16%）,and 68 strains of clustering Escherichia coli（3.68%）.[Conclusion]Taq Man real-time fluorescence PCR combined with the traditional separation method is accurate and efficient in detection of DEC,and can avoid the missing detection of Shigella apps. The monitoring data indicate that DECs are the most common pathogens in the pathogentic spectrum of bacterial infectious diarrhea,and the composition structure shows diverse and significant seasonal characteristics.

关 键 词：致泻性大肠埃希菌 实时荧光PCR 传统分离 EPEC ETEC EIEC EAEC 

分 类 号：R115[医药卫生—公共卫生与预防医学]