CAJ | 학술논문

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.
목적 평고화건립용우상규감측4충치사성대장애희균(DEC)적분자진단방법,병응용우상해시복사인군DEC적감측.방법 사용단맥SSI분자진단시제합대DEC삼고균주진행험증시험급제정DEC-PCR진단、분리적조작규정(DEC-PCR-SOP),병검측2012년6-9월상해시3가림상의원복사병례분편표본.결과 경26주DEC삼비균주험증,SSI분자진단시제합적특이성위100％; 1887빈복사병례표본공분리득도218주DEC(유당양성181주,유당음성37주),기중치병성대장애희균(EPEC) 118주、산독성대장애희균(ETEC)90주、침습성대장애희균(EIEC)9주、산지하독소대장애희균(STEC)1주、지하균18주,총양성솔위l1.6％；감측지구DEC복사병례중이EPEC점우세,이EPEC복사병례중우이2세이하영유인위주；외적DEC병례이ETEC점우세,신생인ETEC병례점5세이하저년령조복사병례적1/3.결론 경평고DEC-PCR-SOP용우4충DEC상규감측적수거결과가신.국내식원성감측망락실험실응불단완선4충DEC진단화삼비능력.
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.