Background and Objectives Transfer of antibiotic resistance genes by integrons is the main cause of drug-resistant bacteria. This study aims to evaluate the prevalence of class I, II, and III integrons among uropathogenic Escherichia Coli (UPEC) strains isolated from patients with Urinary tract infection (UTI). Subjects and Methods This cross-sectional study was carried out on 50 UPEC strains isolated from patients with UTI referred to hospitals in Shiraz, Iran in 2020. Antibiotic susceptibility pattern was evaluated by the disk diffusion susceptibility test. Then, the prevalence of class 1 to 3 integrons in the isolates was investigated by the polymerase chain reaction test. Data were statistically analyzed in SPSS software using chi-square test. P≤0.05 was statistically significant. Results 42% of isolates had multi-drug resistance. The highest antibiotic resistance and sensitivity were related to trimethoprim/sulfamethoxazole (52%) and gentamicin (90%), respectively. There was a significant relationship between the presence of class I integron and resistance to amikacin and ciprofloxacin, between the presence of class II integron and resistance to gentamicin, and between the presence of class III integron and resistance to trimethoprim/sulfamethoxazole and nalidixic acid (P<0.05). Conclusion There is a significant association between the presence of class I, II and III integrons and antibiotic resistance in UPEC strains isolated from patients with UTI. Infection control measures and suitable treatment methods are needed for preventing the spread of these isolates in the hospitals and health centers in Shiraz city.