An outbreak due to Shiga toxin-producing Escherichia coli O26[ratio ]H11 (STEC) occurred at a nursery in southeastern Japan in 1997. Thirty-two children had watery or bloody diarrhoea but none of them suffered from haemolytic-uremic syndrome. All of the STEC O26 were isolated during the period from 23 July to 22 August from 24 children, 3 nurses, and 2 food samples. These organisms had stx1 and eae genes but none of the other genes for which we tested (stx2, bfp, and EAF plasmid). They also possessed multiple antimicrobial resistances, which were encoded by a transmissible plasmid, and showed mostly identical genomic pulsed-field gel electrophoretic patterns. The results of this investigation suggested that contaminated food was the main contributing factor to this multiple antimicrobial-resistant STEC O26 infection, and person-to-person transmission also contributed to the spread of this outbreak.