From a clinical prospective, it is important to know the distribution of virulence factors and antimicrobial resistance properties of Uropathogenic Escherichia coli isolated from diabetic patients with severe urinary tract infections. Therefore, the present study was carried out in order to reach above goals.  Totally, 110 urine samples were collected from diabetic patients hospitalized due to UTIs. Samples were cultured and those that were E. coli positive were subjected to PCR assays to study the distribution of virulence factors. Simple disk diffusion was carried out to study the antibiotic resistance pattern of E. coli isolates.  We found that 33 out of 110 diabetic urine samples (30%) were positive for E. coli. E. coli strains of diabetic patients had the highest levels of resistance to tetracycline (87.8%), ampicillin (84.8%), gentamycin (84.8%), ciprofloxacin (75.7%) and cephalothin (54.5%). In the other hand, imipenem and mezlocillin were the best choices for treatment of the cases of UTIs in diabetic patients. Sfa (45.4%), fim (45.4%) and afa (36.3%) had the highest incidence among virulence factors while, usp (3.0%), sigA (6.0%) and iha (6.0%) had the lowest incidence. Conclusions: Virulent strains of uropathogenic E. coli were predominant cause of UTIs in diabetic patients of Iran. High hygienic care, treatment of diabetes and using from disk diffusion method can reduce the dangerous risk of uropathogenic E. coli in diabetic patients.