Background & aim: About 40% of women and 12% of men experience a urinary tract infection at least once in their lifetime. Aminoglycosides are very effective antimicrobial agents. The emergence and spread of antibiotic resistance in Escherichia coli (E. coli) is becoming a global public health concern. The aim of this study was to determine the presence of ant(2'')-Ia and ant(4')-IIa genes in E. coli strains isolated from urinary tract infections.   Methods: This cross-sectional study was performed on 187 strains of Escherichia coli isolated from urine specimens of patients referred to Amir Al-Momenin Ali Hospital in Gerash city (Fars province) from June 2020 to June 2021. The pattern of antibiotic resistance to tobramycin, gentamicin, kanamycin and amikacin by disk diffusion method was investigated. PCR method was used to evaluate the presence of ant(2'')-Ia and ant(4')-IIa genes. Chi-square test was used for data analysis.   Result: Out of 187 strains studied, 62 strains (33.15%) were resistant to aminoglycosides. The highest and lowest aminoglycoside resistance was observed against tobramycin (79.03%) and amikacin (11.29%), respectively. Two strains (3.22%) were resistant to all aminoglycosides. ant(2'')-Ia and ant(4')-IIa genes were identified in 17 strains (27.41%) and 11 strains (17.74%), respectively.   Conclusion: The results of the present study show that Escherichia coli strains isolated from urinary tract infections have relatively low resistance to common aminoglycosides. For the initial treatment of urinary tract infections caused by Escherichia coli in Gerash city, the use of amikacin and kanamycin antibiotics is recommended.