| Summary: | <i>Escherichia coli</i> causing urinary tract infections (UTIs) are one of the most common outpatient bacterial infections. This study aimed to compare the characteristics of <i>E. coli</i> isolated from UTI patients in a single medical center in 2009–2010 (n = 504) and 2020 (n = 340). The antimicrobial susceptibility of <i>E. coli</i> was determined by the disk diffusion method. PCRs were conducted to detect phylogenetic groups, ST131, K1 capsule antigen, and 15 virulence factors. Phylogenetic group B2 dominated in our 2009–2010 and 2020 isolates. Moreover, no phylogenetic group E strains were isolated in 2020. <i>E. coli</i> isolates in 2020 were more susceptible to amoxicillin, ampicillin/sulbactam, cefuroxime, cefmetazole, ceftazidime, cefoxitin, tetracycline, and sulfamethoxazole/trimethoprim, compared to the isolates in 2009–2010. Extensively drug-resistant (XDR)-<i>E. coli</i> in 2009–2010 were detected in groups B1 (5 isolates), B2 (12 isolates), F (8 isolates), and unknown (1 isolate). In 2020, XDR-<i>E. coli</i> were only detected in groups A (2 isolates), B2 (5 isolates), D (1 isolate), and F (4 isolates). The prevalence of virulence factor genes <i>aer</i> and <i>fimH</i> were higher in <i>E. coli</i> in 2009–2010 compared to those in 2020. In contrast, <i>afa</i> and <i>sat</i> showed higher frequencies in <i>E. coli</i> isolates in 2020 compared to <i>E. coli</i> in 2009–2010.
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