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Journal of Clinical Microbiology, January 2009, p. 189-197, Vol. 47, No. 1
0095-1137/09/$08.00+0     doi:10.1128/JCM.01282-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Phenotypic and Genotypic Analysis of Enterotoxigenic Escherichia coli in Samples Obtained from Egyptian Children Presenting to Referral Hospitals{triangledown}

H. I. Shaheen,1* I. A. Abdel Messih,1 J. D. Klena,1 A. Mansour,1 Z. El-Wakkeel,2 T. F. Wierzba,1,{dagger} J. W. Sanders,1,{ddagger} S. B. Khalil,1 D. M. Rockabrand,1,§ M. R. Monteville,1 P. J. Rozmajzl,1,¶ A. M. Svennerholm,3 and R. W. Frenck1,||

U.S. Naval Medical Research Unit No. 3, Cairo, Egypt,1 Ministry of Health and Population, Cairo, Egypt,2 Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden3

Received 8 July 2008/ Returned for modification 3 October 2008/ Accepted 16 October 2008

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.

* Corresponding author. Mailing address: U.S. Naval Medical Research Unit No. 3, Cairo, Egypt. Phone: 202 23480324. Fax: 202 23483482. E-mail: hind.shaheen.eg{at}med.navy.mil

{triangledown} Published ahead of print on 29 October 2008.

{dagger} Present address: U.S. Naval Medical Research Unit Number 2, Cambodia.

{ddagger} Present address: U.S. Naval Medical Research Center Detachment, Unit 3800, APO, Lima, Peru.

§ Present address: U.S. Naval Medical Research Center, Silver Spring, MD 20910.

Present address: Navy Environmental and Preventive Medicine Unit 6, 1250 North Road Pearl Harbor, Pearl Harbor, HI 96860.

|| Present address: Cincinnati Children's Hospital Medical Center, 3333 Burnet Ave., MLC 6014, Cincinnati, OH 45229.

Journal of Clinical Microbiology, January 2009, p. 189-197, Vol. 47, No. 1
0095-1137/09/$08.00+0     doi:10.1128/JCM.01282-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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