We examined the prevalence and characteristics of extended-spectrum beta-lactamase (ESBL)-producing clinical isolates among Enterobacter spp., Serratia marcescens, Citrobacter freundii, and Morganella morganii, and evaluated screening criteria, clinical characteristics and outcomes of infections caused by ESBL-producing organisms. Between January and June 2005, a total of 493 nonduplicate consecutive isolates were collected at Asan Medical Center, a 2,300-bed tertiary hospital in Seoul, Republic of Korea. Fifty isolates (10.1%) were positive for phenotypical ESBL-test. The positive rate of phenotypical ESBL-test in Enterobacter spp., S. marcescens, C. freundii, and M. morganii was 12.8%, 12.4%, 4.9%, and 0% respectively. SHV-12 (18 isolates), CTX-M-9 (17 isolates), and TEM-52 (five isolates) were the most prevalent ESBL types. The ESBL in 17 strains could not be identified. As an ESBL screening criterion, the cefepime MIC >or=1 microg/ml had the highest sensitivity (0.84) and specificity (0.87). Half of the ESBL-producing isolates (25/50) were judged as pathogens. Cholangitis (ten cases), and pneumonia (six cases) were the most common infections. The overall mortality was 12.0%.