Diarrheagenic Escherichia coli (DEC) is an emerging agent among pathogens that causes diarrhea. Studies showed that diarrheagenic E. coli such as enterohaemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC), diffusely adhering E. coli (DAEC) and shiga toxin producing E. coli (STEC) strains are among the most frequent causative agents in acute diarrhea. The aim of this study was to determine the frequency of DEC pathotypes in Khuzestan province.Stool samples were collected from patients with diarrhea in Khuzestan province of Iran. E. coli strains were isolated using conventional culture and standard biochemical tests. The polymerase chain reaction (PCR) technique was used to detect presence of virulence genes, i.e; eae, stx1 and stx2 for EHEC, bfp and eae for EPEC, LT and ST for ETEC, AA for EAEC, invE for EIEC, stx1 and stx2 for STEC.Altogether, 200 stool samples were obtained from patients, of which 158 (79%) were positive for E. coli. DEC was identified in 127 (63%) of stool samples, which frequency of each pathotypes were as follows: atypical EPEC 49 (39%), typical EPEC 1 (0.7%), STEC 50 (39.3%), ETEC 21 (16.3%), EAEC 5 (4.0%) and EIEC 1 (0.7%). Most frequent etiological agents of diarrhea in Khuzestan province of Iran were STEC and EPEC.Our findings showed DEC had been agent of diarrhea in Khuzestan. This finding provides evidence that effort should be made to estimate the burden of infection by the etiological agent for better medical approach and should raise notification about antibiotic resistance among bacterial infection.