Identification, Characterization, and Distribution of a Shiga Toxin 1 Gene Variant ( stx 1c ) in Escherichia coli Strains Isolated from Humans

Abstract

ABSTRACT By using sequence analysis of Shiga toxin 1 (Stx 1) genes from human and ovine Stx-producing Escherichia coli (STEC) strains, we identified an Stx1 variant in STEC of human origin that was identical to the Stx1 variant from ovine STEC, but demonstrated only 97.1 and 96.6% amino acid sequence identity in its A and B subunits, respectively, to the Stx1 encoded by bacteriophage 933J. We designated this variant “Stx1c” and developed stxB 1 restriction fragment length polymorphism and stx 1c -specific PCR strategies to determine the frequency and distribution of stx 1c among 212 STEC strains isolated from humans. stx 1c was identified in 36 (17.0%) of 212 STEC strains, 19 of which originated from asymptomatic subjects and 16 of which were from patients with uncomplicated diarrhea. stx 1c was most frequently (in 23 STEC strains [63.9%]) associated with stx 2d , but 12 (33.3%) of the 36 STEC strains possessed stx 1c only. A single STEC strain possessed stx 1c together with stx 2 and was isolated from a patient with hemolytic-uremic syndrome. All 36 stx 1c -positive STEC strains were eae negative and belonged to 10 different serogroups, none of which was O157, O26, O103, O111, or O145. Stx1c was produced by all stx 1c -containing STEC strains, but reacted weakly with a commercial immunoassay. We conclude that STEC strains harboring the stx 1c variant account for a significant proportion of human STEC isolates. The procedures developed in this study now allow the determination of the frequency of STEC strains harboring stx 1c among clinical STEC isolates and their association with human disease in prospective studies.