Background: Urinary tract infections are among the most common bacterial infections in clinical practice. The strains of Extended-spectrum beta-lactamase (ESBL) producing Escherichia coli, represent serious therapy problem and significant concern. Objectives and Methods: To determine the presence and antibiotic resistance pattern of ESBL producing E. coli in urine samples in the period of 3 years (2021-2023). Urine samples sent in the Department of Microbiology at the Center for Public Health Bitola, were inoculated on Chromogenic media (CPS agar, bioMerieux). Double-disk synergy (DDS) test and Automatized Vitek 2 system were used for detection of ESBL-producing strains. Antimicrobial susceptibility was examined by standard disk diffusion method and MIC concentrations with Automatized Vitek 2 system. Results: Out of the total number of E. coli 1201 in 2021, 1443 in 2022 and 1776 in 2023, ESBL positive were 56 (4.66%), 134 (9.28%) and 90 (5.06%) respectively. All 280 ESBL-producing isolates, showed high percentage of resistance against all cephalosporins as follows: 100% to cefuroxime, 100% to ceftriaxone, 86.34% to ceftazidime and 69.91% to cefepime. While low level of resistance was observed against fosfomycin (2.34%) and nitrofurantoin (4.39%), and no resistance to carbapenems. Conclusions: The results indicate the need of constant monitoring of the percentage of antibiotic resistance of uropathogen bacteria in order to make the correct choice of effective antibiotics which rapidly and efficiently will treat and prevent recurrent urinary infections.