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从埃及转诊医院就诊儿童样本中分离的产肠毒素大肠杆菌的表型和基因型分析

Phenotypic and genotypic analysis of enterotoxigenic Escherichia coli in samples obtained from Egyptian children presenting to referral hospitals.

作者信息

Shaheen H I, Abdel Messih I A, Klena J D, Mansour A, El-Wakkeel Z, Wierzba T F, Sanders J W, Khalil S B, Rockabrand D M, Monteville M R, Rozmajzl P J, Svennerholm A M, Frenck R W

机构信息

U.S. Naval Medical Research Unit No. 3, Cairo, Egypt.

出版信息

J Clin Microbiol. 2009 Jan;47(1):189-97. doi: 10.1128/JCM.01282-08. Epub 2008 Oct 29.

Abstract

Hospital surveillance was established in the Nile River Delta to increase the understanding of the epidemiology of diarrheal disease among Egyptian children. Between September 2000 and August 2003, samples obtained from children less than 5 years of age who had diarrhea and who were seeking hospital care were cultured for enteric bacteria. Colonies from each culture with a morphology typical of that of Escherichia coli were tested for the heat-labile (LT) and heat-stable (ST) toxins by a GM-1-specific enzyme-linked immunosorbent assay and colonization factor (CF) antigens by an immunodot blot assay. Enterotoxigenic E. coli (ETEC) isolates were recovered from 320/1,540 (20.7%) children, and ETEC isolates expressing a known CF were identified in 151/320 (47%) samples. ST CFA/I, ST CS6, ST CS14, and LT and ST CS5 plus CS6 represented 75% of the CFs expressed by ETEC isolates expressing a detectable CF. Year-to-year variability in the proportion of ETEC isolates that expressed a detectable CF was observed (e.g., the proportion that expressed CFA/I ranged from 10% in year 1 to 21% in year 3); however, the relative proportions of ETEC isolates expressing a CF were similar over the reporting period. The proportion of CF-positive ETEC isolates was higher among isolates that expressed ST. ETEC isolates expressing CS6 were isolated significantly less often (P < 0.001) than isolates expressing CFA/I in children less than 1 year of age. Macrorestriction profiling of CFA/I-expressing ETEC isolates by using the restriction enzyme XbaI and pulsed-field gel electrophoresis demonstrated a wide genetic diversity among the isolates that did not directly correlate with the virulence of the pathogen. The genome plasticity demonstrated in the ETEC isolates collected in this work suggests an additional challenge to the development of a globally effective vaccine for ETEC.

摘要

在尼罗河三角洲建立了医院监测体系,以增进对埃及儿童腹泻病流行病学的了解。2000年9月至2003年8月期间,对因腹泻而寻求医院治疗的5岁以下儿童的样本进行肠道细菌培养。对每种培养物中形态典型的大肠杆菌菌落,通过GM-1特异性酶联免疫吸附测定法检测热不稳定(LT)和热稳定(ST)毒素,并通过免疫斑点印迹测定法检测定植因子(CF)抗原。从1540名儿童中的320名(20.7%)分离出了产肠毒素大肠杆菌(ETEC),在320份样本中的151份(47%)中鉴定出表达已知CF的ETEC分离株。ST CFA/I、ST CS6、ST CS14以及LT和ST CS5加CS6占表达可检测CF的ETEC分离株所表达CF的75%。观察到表达可检测CF的ETEC分离株比例存在逐年变化(例如,表达CFA/I的比例从第1年的10%到第3年的21%);然而,在报告期内表达CF的ETEC分离株的相对比例相似。表达ST的分离株中CF阳性ETEC分离株的比例更高。在1岁以下儿童中,表达CS6的ETEC分离株的分离频率明显低于表达CFA/I的分离株(P<0.001)。使用限制性内切酶XbaI和脉冲场凝胶电泳对表达CFA/I的ETEC分离株进行宏观限制性图谱分析,结果表明这些分离株之间存在广泛的遗传多样性,且与病原体的毒力没有直接关联。在这项研究中收集的ETEC分离株中显示出的基因组可塑性表明,开发一种全球有效的ETEC疫苗面临额外挑战。

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