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Uropathogenic Escherichia coli (UPEC) is a well-known pathogen that has perturbed the medical scenario because of its resistance to diverse therapeutic drugs and its ability to form a biofilm. Different O-serogroups are the prevalent cause of urinary tract infections (UTIs) along with their ability to form a biofilm. The present research aimed to assess antibiotic susceptibility, biofilm formation, and serotyping of UPEC isolates in conjunction with the demographic data. Antibiotic susceptibility was determined using the Kirby–Bauer method and biofilm formation was assessed phenotypically and at the molecular level. Serotyping was performed by multiplex PCR. A significant proportion of the total of 120 UPECs was isolated from women (p < 0.05). Most isolates were resistant to cefotaxime, ceftazidime, and tetracycline, but maintained their sensitivity to imipenem. O₂₅, O₁₅, O₈, and O₇₅ were the most commonly …