To describe the epidemiology of Enterobacteriaceae-producing extended-spectrum beta-lactamase (EP-ESBL) in a non-outbreak setting, and to define the risk factors associated with colonization, a 5-month surveillance study was initiated. Ten of 333 patients were colonized with EP-ESBL, as defined by isoelectric focusing, Klebsiella sp. and Escherichia coli were the species most commonly harbouring these plasmid-mediated enzymes. Of the 16 SHV-producing isolates, 10 were SHV-3-like (pI 7.0) and six were SHV-like (pI 8.2). All isolates were resistant to ceftriaxone. Ceftazidime resistance was detected in 50% and 100% of SHV-3-like and SHV-5-like producing isolates, respectively. One patient was colonized with four different SHV-5-like producing Enterobacteriaceae. These isolates carried plasmids that were indistinguishable by restriction endonuclease analysis, indicating broad plasmid transfer within the patient. By logistic regression, haemodialysis was a strong risk factor for colonization with EP-ESBL, suggesting that, in our hospital, horizontal transmission is an important mechanism of dissemination of these resistant pathogens.