Enteropathogenic Escherichia coli (EPEC) strains possess genes for attaching and effacing (eae) and EPEC adherence factor (EAF) plasmid. It is necessary to develop molecular techniques for the evaluation of EPEC isolates. A total of 183 E. coli isolates from neonates admitted to Pusan National University Hospital were investigated by polymerase chain reaction (PCR) and DNA colony hybridization. Of the 183 isolates tested, 10 (5.5%) were positive for eae by PCR and DNA colony hybridization and confirmed to be EPEC. Ten EPEC isolates showed 3 different adherence patterns: seven strains had diffuse adherence, two localized adherence-like adherence, and one aggregative adherence. They were also examined by antimicrobial susceptibility tests, serotyping, and molecular epidemiological typing such as pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis. The EPEC isolates could be divided into 9 different antimicrobial resistance patterns, 6 serotypes, 4 PFGE patterns, and 5 RAPD patterns. This result indicates that the EPEC isolates from neonates were originated from different sources.