4种致泻性大肠埃希菌分子诊断方法的评估及其在监测中的应用

黄峥; 许浩; 郭家胤; 黄晓兰; 李颖; 侯琪; 王爱敏; 王传清; 金汇明; 许学斌; 胡家瑜; 史贤明; 冉陆; 阚飚

Abstract

黄峥,许浩,郭家胤,黄晓兰,李颖,侯琪,王爱敏,王传清,金汇明,许学斌,胡家瑜,史贤明,冉陆,阚飚.4种致泻性大肠埃希菌分子诊断方法的评估及其在监测中的应用[J].Chinese journal of Epidemiology,2013,34(6):614-617

4种致泻性大肠埃希菌分子诊断方法的评估及其在监测中的应用

Assessment and application of a molecular diagnostic method on the detection of four types of diarrheagenic Escherichia coli

Received:January 14, 2013 Revised:August 15, 2012

DOI：

KeyWord: 致泻性大肠埃希菌分子诊断评估监测

English Key Word: Diarrheagenic Escherichia coli Molecular diagnostics Assessment Monitor

FundProject:国家重大科技专项;中美新发再发传染病合作项目;国家高技术研究发展计划("863"计划)

Author Name	Affiliation	E-mail
HUANG Zheng	Center of Hygiene Inspection of Changning District , Shanghai 200051,China	xbxu@scdc.sh.cn;chuangqing12@yahoo.com.cn
XV Hao	Center of Hygiene Inspection of Changning District , Shanghai 200051,China
GUO Jia-yin	Center of Hygiene Inspection of Changning District , Shanghai 200051,China
HUANG Xiao-lan	Changning District Center of Disease Control and Prevention
LI Ying	Changning District Center of Disease Control and Prevention
HOU Qi	Shanghai United Family Hospital
WANG Ai-min	Fu-Dan University Subsidiary Children Hospital
WANG Chuan-qing	Fu-Dan University Subsidiary Children Hospital
JIN Hui-ming	Shanghai Municipal Center of Disease Control and Prevention
XU Xue-bin	Shanghai Municipal Center of Disease Control and Prevention
HU Jia-yu	Shanghai Municipal Center of Disease Control and Prevention
SHI Xian-ming	School of Agriculture and Biology, Shanghai Jiaotong University
RAN Lu	National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention
KAN Biao	National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention

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Abstract:

目的评估和建立用于常规监测4种致泻性大肠埃希菌(DEC)的分子诊断方法,并应用于上海市腹泻人群DEC的监测.方法使用丹麦SSI分子诊断试剂盒对DEC参考菌株进行验证试验及制定DEC-PCR诊断、分离的操作规程(DEC-PCR-SOP),并检测2012年6-9月上海市3家临床医院腹泻病例粪便标本.结果经26株DEC参比菌株验证,SSI分子诊断试剂盒的特异性为100%; 1887份腹泻病例标本共分离得到218株DEC(乳糖阳性181株,乳糖阴性37株),其中致病性大肠埃希菌(EPEC) 118株、产毒性大肠埃希菌(ETEC)90株、侵袭性大肠埃希菌(EIEC)9株、产志贺毒素大肠埃希菌(STEC)1株、志贺菌18株,总阳性率为l1.6%；监测地区DEC腹泻病例中以EPEC占优势,而EPEC腹泻病例中又以2岁以下婴幼儿为主；外籍DEC病例以ETEC占优势,新生儿ETEC病例占5岁以下低年龄组腹泻病例的1/3.结论经评估DEC-PCR-SOP用于4种DEC常规监测的数据结果可信.国内食源性监测网络实验室应不断完善4种DEC诊断和参比能力.

English Abstract:

Objective To establish and evaluate a molecular diagnostic method for routine monitoring of four types of diarrheagenic Escherichia (E.) coli (DEC) and to study the distribution of four types of DEC isolated from diarrheal patients in Shanghai.Methods DEC-PCR standard operation procedure (SOP) had been developed for DEC detection and isolation,using the Statens Serum Institute (SSI) DEC PCR kits with multiplex PCR technique after verification tests on reference strains.Diarrhea specimens from 3 clinical hospitals in Shanghai were tested from June to September,2012.Results Specificity of the PCR kit was 100% by verification on the 26 DEC reference strains.A total number of 218 DEC isolates,including 181 fermented lactose and 37 unfermented lactose were identified from the 1887 stool specimens of diarrhea patients,with positive rate as 11.6%.The most common pathogen (54.1%,118/218) was enteropathogenic E.coli (EPEC),followed by enterotoxigenic E.coli (ETEC,41.3%,90/218),enteroinvasive E.coli (EIEC,4.1%,9/218) and Shigatoxin-producing E.coli (STEC,0.5%,1/218) in addition to 18 Shigella isolates.ETEC dominated in diarrhea patients with foreign residency,as well as 1/3 were perinatal stage of neonatal ETEC of all diarrhea cases under the age of 5,while EPEC dominated in the Chinese diarrhea patients especially among young kids under the age of 2.Conclusion Data was reliable after assessment on this molecular diagnostics and seperation procedures used for the routine monitoring on four types of DEC,while the diagnosis and reference ability of DEC regarding the laboratories net-working on food-borne pathogens need to be built up and improved.

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