Hemorrhagic Escherichia coli (E. coli) is one of the most important agent of diarrhea in piglet. The aim of this study was to investigate the characteristics of virulence genes and genetic diversity in hemorrhagic E. coli isolated from piglets with diarrhea. Among 122 hemorrhagic E. coli, 62 isolates carried single toxin gene like heat-stable toxin (ST), heatlabile toxin (LT), verotoxin 1 (VT1), verotoxin 2 (VT2), spa and eae. The most prevalent toxin gene carried by isolates was ST gene (23 isolates), while the most common association was ST/LT (14 isolates) and ST/LT/VT2 (13 isolates). In pulsed field gel electrophoresis, isolates were not classified as one cluster by epidemiological information including isolated area, isolated year and possessed toxins.