Our objective was to analyse phenotypic and genetic data of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae, Enterobacter cloacae, Escherichia coli and Serratia marcescens that cause infections in our hospital. Over a 3 year period, 342 randomly selected clinical Enterobacteriaceae isolates were tested for ESBL production and evaluated for the presence of the β-lactamase genes bla(SHV), bla(TEM,) bla(CTX-M) and bla(TLA-1). The antibiotic susceptibilities of these isolates were also determined, and the clonality of the isolates was assessed by PFGE. Based on our analyses, 33/92 (35.9 %) K. pneumoniae, 31/87 (35.6 %) Enterobacter cloacae, 24/80 (30 %) E. coli and 17/83 (20.5 %) S. marcescens were identified as ESBL producers. The presence of TEM, SHV or CTX ESBL types was detected in 99/105 (94 %) of the isolates. TLA-1 was not detected in any of the 105 isolates. The dominant ESBL types were bla(SHV-5) (n=33), bla(SHV12) (n=31) and bla(CTX-M-15) (n=30). The predominant ESBL identified in E. coli and Enterobacter cloacae isolates was CTX-M-15, whereas in K. pneumoniae and S. marcescens the predominant types were SHV-12 and SHV-5, respectively. PFGE genotyping revealed two main genetic patterns in the K. pneumoniae isolates, types SHV-12 and TEM-1+SHV-5. An outbreak caused by Enterobacter cloacae SHV-5+CTX-M-15 was detected. In contrast, most ESBL-producing isolates of E. coli and S. marcescens did not have similar PFGE banding patterns and thus were not genetically similar. Enterobacteriaceae are a concern in our hospital, especially K. pneumoniae and Enterobacter cloacae. Our results confirm that the CTX-M-15 ESBL type has spread rapidly in the hospital, and thus requires careful monitoring.