We have assessed the frequency of Shiga toxin-producing Escherichia coil (STEC) in clinical and food samples as well as studied the genotypic and phenotypic characteristics of the recovered strains. One hundred ninety eight fecal samples from children with bloody diarrhea (BD), 14 from children with hemolytic uremic syndrome (HUS), 220 ground beef samples and 4 STEC isolates from other beef-derived products were analyzed. The STEC strains were isolated from 3 (1.5%) children with bloody diarrhea, 1 (7%) from a child with HUS and 4 (1.8%) from ground beef samples. All strains were eae and ehxA positive. The serotypes found were: O157:H7 (9 strains), O26:H11 (2), O111: NM (1) and O145:HNT (1). All O157:H7 STEC strains harbored the eae subtype gamma1, O26:H11 and O145:HNT strains, subtype beta1 and O111:NM strain, subtype gamma2/theta. The STEC strains of the same serogroup showed high genetic diversity. In Uruguay, STEC is not frequently isolated from cases of bloody diarrhea in children. However, all the recovered STEC strains carried the genes associated with severe disease and 2 out of 3 children infected with STEC developed HUS. Ground beef and other food products might be important vehicles for O157:H7 strains.