Background: Esherichia coli is a significant species of bacteria connected with nosocomial infections involving, pneumonia, infections of urinary tract and damage of the surgical site. Aim of the Study: The aim of this thesis is to investigate the occurrence of genes encoding AMEs enzymes in aminoglycosides resistance E.coli isolates recovered from urinary tract infection in Najaf hospitals. Methods: A total of 68 specimens were isolated from patients with UTI at burn cente in Najaf City from the whole study period. The 24 isolates were screened for the sensitivity versus 3 various antibiotics disks of the one class by using Kirby-Bauer disk diffusion method. Results: the resistance profiles of the isolates show that 21 (91.3%) were resistant to 1 or more antibiotics of aminoglycoside. Present study show that 20 (86.95%) of XDR isolates harbored aac(6')-Ib gene. Conclusion: The study emphasizes the significance of aminoglycoside resistant E.coli isolates as a cause of UTI infection in Najaf City.