Urinary tract infections are common and occasionally life threatening condition amongst diabetic and non-diabetic patients. The present study investigated the distribution of β-lactamase enzyme producing uropathogenic E.coli isolates amongst diabetic and non-diabetic patients. Collected isolates were identified and confirmed as uropathogenic E.coli by standard microbiological procedure and antibiotic susceptibility were screened by Kirby-Bauer disc diffusion method. Phenotypic detection of β- lactamases such as ESBL, AmpC β- lactamase and carbapenemase were determined by double disc diffusion method, disc approximation test and modified Hodge test respectively. β- lactamase encoding genes such as TEM, SHV,CTX-M for ESBLs, ACC, EBC, CIT,DHA, MOX and FOX for AmpC-β-lactamase and KPC, IMP, VIM, NDM and OXA-48 for carbapenemase were detected by PCR method. Out of 306 isolates, 280 (140 from diabetic and 140 from non-diabetic UTI patients) non-repetitive isolates were identified and confirmed as Uropathogenic E.coli. Antibiotic screening revealed that 124 diabetic (88.57%) and 117 non-diabetic (83%) isolates were resistant to at least one antibiotic included in the study. Phenotypic confirmation of various β-lactamase enzymes such as ESBL (diabetic=85/85; 100% and non-diabetic=78/79; 98.7%), AmpC β-lactamase (diabetic=26/36; 72.2% and non-diabetic=18/26; 69%) and carbapenemase (diabetic=19/25; 76% and non-diabetic=13/15; 86.6%) were determined. ESBL encoding genes (diabetic=74/85; 87.05% and non-diabetic=65/78(83.3%), AmpC β- lactamase encoding genes (diabetic=21/26; 80.8% and non-diabetic=12/18; 66.6%) and carbapenemase encoding genes (diabetic=18/19; 94.73% and non-diabetic=11/13; 84.61%) were genotypically confirmed. This study showed that the number of beta-lactamase producing UPEC isolates were phenotypically and genotypically higher in diabetic than non-diabetic patients.