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Journal of Clinical Microbiology, November 2003, p. 5022-5032, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5022-5032.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Distribution of Intimin Subtypes among Escherichia coli Isolates from Ruminant and Human Sources

Elizabeth Macarthur Agricultural Institute, Camden, New South Wales, 2570,¹ Department of Biological Sciences, University of Wollongong, New South Wales, 2522,² Microbiological Diagnostic Unit, Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria, 3052, Australia³

Received 2 January 2003/ Returned for modification 9 February 2003/ Accepted 22 August 2003

The intimin gene eae, located within the locus of enterocyte effacement pathogenicity island, distinguishes enteropathogenic Escherichia coli (EPEC) and some Shiga toxin-producing E. coli (STEC) strains from all other pathotypes of diarrheagenic E. coli. EPEC is a leading cause of infantile diarrhea in developing countries, and intimin-positive STEC isolates are typically associated with life-threatening diseases such as hemolytic-uremic syndrome and hemorrhagic colitis. Here we describe the development of a PCR-restriction fragment length polymorphism (RFLP) assay that reliably differentiates all 11 known intimin types (1, 2, ß, , , , , , , , and ) and three new intimin genes that show less than 95% nucleotide sequence identity with existing intimin types. We designated these new intimin genes Int-µ, Int-, and Int-. The PCR-RFLP assay was used to screen 213 eae-positive E. coli isolates derived from ovine, bovine, and human sources comprising 60 serotypes. Of these, 82 were STEC isolates, 89 were stx-negative (stx^-) and ehxA-positive (ehxA⁺) isolates, and 42 were stx^- and ehxA-negative isolates. Int-ß, the most commonly identified eae subtype (82 of 213 [38.5%] isolates), was associated with 21 serotypes, followed by Int- (39 of 213 [18.3%] isolates; 11 serotypes), Int- (25 of 213 [11.7%] isolates; 15 serotypes), Int- (19 of 213 [8.9%] isolates; 9 serotypes), and Int- (21 of 213 [9.9%] isolates; 5 serotypes). Intimin subtypes 1, 2, , , , µ, , and were infrequently identified; and Int- was not detected. Phylogenetic analyses with the Phylip package of programs clustered the intimin subtypes into nine distinct families (, ß-, , , --, -µ, , , and ). Our data confirm that ruminants are an important source of serologically and genetically diverse intimin-containing E. coli strains.

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