Background: The increased rate of urinary tract infection (UTI) in immunocompromised patients especially diabetic patients is a major public health problem in adults. Moreover, the infection with multidrug resistant strains producing extended spectrum beta-lactamase (ESBL) is a key obstacle in disease management among such vulnerable population. An immediate proper treatment depends on rapid diagnosis of UTI and screening of antimicrobial resistant pattern with highly sensitive methods which also reduces the possible urinary complications among the diabetic patients. Hence, this study was aimed to determine the occurrence of antibiotic resistant genes for β-lactamases; blTEM and blaCTX-M in uropathogenic Escherichia coli isolates from UTI suspected diabetic and non-diabetic patients. attendingMethods: A hospital-based cross-sectional study was conducted in Kathmandu Model Hospital in association with Central Department of Microbiology, TU from June to December 2018. A total of 1267 non-duplicate mid-stream urine specimens from diabetic and non-diabetic patients were obtained and processed immediately for isolation of uropathogens. The isolates were subjected for antibiotic susceptibility testing and ESBL confirmation. Finally, blaTEM and blaCTX-M ESBL genes were screened by using specific primers.Results: The overall prevalence of the urinary tract infection UTI was found to be 17.20%(218/1267) , out of which diabetic patients were significantly more infected with UTI accounting for 32.29%(31/96) as compared to non-diabetic persons, 15.97%(187/1171). A total of 221 bacterial were from 218 culture positive specimens in which E . coli was a most predominate one; 67.9%(150/221.). Forty-four percent (66/150) of the total E. coli was MDR and 37.33%(56/150) were ESBL producers. Among 56 isolates, 92.3%(12/13) from diabetic patents and 83.0% (44/53) were from non-diabetic patients. Furthermore, 84.85% of the screened ESBL producers were confirmed to possess either single or both of the blaTEM and blaCTX-M genes . The blaTEM and blaCTX-M genes were detected in 53.57% and 87.5% of the phenotypically ESBL confirmed E. coli .Conclusions: The UTI infection is an increasing problem in diabetic patients and infection with multidrug resistant strains specially ESBL producing uropathogens are causing a huge problem in disease management leading to high rate of mortality and morbidity of diabetic patients.