<b>Background and Objective:</b> Urinary tract infections believe to be one of the main acquainted infections by <i>Escherichia coli</i> in hospitals with an excessive incidence of illness. This study aimed to analyze the antibiotic resistance profile and molecular characteristics of <i>E. coli</i> isolates recovered from patients with urinary tract infection at different hospitals in Taif Governorate, Saudi Arabia. <b>Materials and Methods:</b> Out of 143 isolates collected for 11 months, from February-December 2019, 24 isolates were identified as <i>E. coli</i> by API system and 16S rRNA sequences techniques. An antibiotic sensitivity test was performed using the disk diffusion method. Besides, the repetitive sequence repeat-PCR (Rep-PCR) technique was used for genotyping the 24 isolates. <b>Results:</b> Almost all isolates were resistant to most tested antibiotics such as ampicillin, ceftazidime, cefepime, trimethoprim/sulfamethoxazole, amox/clavulanic. The PCR results show that virulence genes <i>kpsII</i> and <i>yaiO</i> were detected in all <i>E. coli</i> isolates. <i>Stx1</i>, <i>fimH</i>, <i>hly</i> and <i>uidA</i> were moderate detected in all isolates. <b>Conclusion:</b> The high frequencies of antibiotic-resistant <i>E. coli</i> isolates in patients with urinary tract infections in the current study suggest that continuous surveillance of the use of appropriate antibiotics is required and that control of infections is necessary.
Keywords: 16S rRNA; E. coli; Molecular characterization; polymorphic; rep-PCR; urinary tract infection; virulence genes.