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Clinical and Diagnostic Laboratory Immunology, January 2002, p. 46-53, Vol. 9, No. 1
1071-412X/01/$04.00+0     DOI: 10.1128/CDLI.9.1.46-53.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Immunological Characterization of Escherichia coli O157:H7 Intimin 1

W.-G. Son, T. A. Graham, and V. P. J. Gannon^*

Population and Public Health Branch, Health Canada, Lethbridge, Alberta T1J 3Z4, Canada

Received 20 March 2001/ Accepted 9 October 2001

Portions of the intimin genes of Escherichia coli O157:H7 strain E319 and of the enteropathogenic E. coli O127:H6 strain E2348/69 were amplified by PCR and cloned into pET-28a(+) expression vectors. The entire 934 amino acids (aa) of E. coli O157:H7 intimin, the C-terminal 306 aa of E. coli O157:H7 intimin, and the C-terminal 311 aa of E. coli O127:H6 intimin were expressed as proteins fused with a six-histidine residue tag (six-His tag) in pET-28a(+). Rabbit antisera raised against the six-His tag-full-length E. coli O157:H7 intimin protein fusion cross-reacted in slot and Western blots with outer membrane protein preparations from the majority of enterohemorrhagic and enteropathogenic E. coli serotypes which have the intimin gene. The E. coli strains tested included isolates from humans and animals which produce intimin types (O serogroups 86, 127, and 142), ß1 (O serogroups 5, 26, 46, 69, 111, 126, and 128), 1 (O serogroups 55, 145, and 157), 2 (O serogroups 111 and 103), and (O serogroup 103) and a nontypeable intimin (O serogroup 80), results based on intimin type-specific PCR assays. Rabbit antisera raised against the E. coli O157:H7 C-terminal fusion protein were much more intimin type-specific than those raised against the full-length intimin fusion protein, but some cross-reaction with other intimin types was also observed for these antisera. In contrast, the monoclonal antibody Int1.C11, raised against the C-terminal E. coli O157 intimin, reacted only with preparations from intimin 1-producing E. coli strains such as E. coli O157:H7.

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* Corresponding author. Mailing address: Population and Public Health Branch, Health Canada, Township Road 9-1, Lethbridge, Alberta T1J 3Z4, Canada. Phone: (403) 382-5514. Fax: (403) 381-1202. E-mail: gannonv{at}em.agr.ca.

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Clinical and Diagnostic Laboratory Immunology, January 2002, p. 46-53, Vol. 9, No. 1
1071-412X/01/$04.00+0     DOI: 10.1128/CDLI.9.1.46-53.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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