Background: It has been interesting to compare the levels of antimicrobial resistance and the virulence characteristics of uropathogenic Escherichia coli (UPEC) strains of certain phylogenetic groups. The purpose of this study was to identify the frequency of phylogenetic groups, adhesin genes, antibiotic sensitivity patterns, and extended spectrum beta-lactamase (ESBL) genes in hospital-acquired UPEC. Methods After UPEC isolation, the disc diffusion method was used to assess its susceptibility to antibiotics. Combination disc testing confirmed the existence of ESBL producers. Polymerase chain reaction (PCR) was used to detect genes for adhesin and ESBLs. Results One hundred and twenty-eight E. coli were isolated, which had the highest resistance to tetracycline (96%), followed by cefoxitin (93%), cefepime (92%), ceftazidime (79%), aztreonam (77%), and sulfamethoxazole-trimethoprim (75%). About 57% of isolates were phenotypically ESBL-positive, and they were confirmed by PCR. B2 phylogroup (41%) was the most frequent in E. coli isolates, then group D (30%), group A (18%), and lastly group B1 (11%). ESBL genes were more significantly prevalent in phylogroups B2 and D than other phylogroups (P < 0.001). Regarding adhesin genes, both fimH and afa were more significantly associated with group B2 than other groups (P < 0.009, < 0.032), respectively. In ESBL-positive isolates, both genes were more significantly detected compared to negative ones (P < 0.001). Conclusion Phylogroups B2 and D of UPEC are important reservoirs of antimicrobial resistance and adhesion genes. Detection of ESBL-producing E. coli is important for appropriate treatment as well as for effective infection control in hospitals.