Background The risk of urinary tract infection (UTI) is higher in diabetics compared to non-diabetics. The etiology and the antibiotic resistance of uropathogens have been changing over the past years. Hence the study was undertaken to determine if there are differences in clinical and microbiological features of UTI between diabetic and non-diabetic subjects, to study the influence of diabetes mellitus on the uropathogens and antibiotic sensitivity pattern in patients with UTI. Method A total of 181 diabetics (83 males and 98 females) and 124 non-diabetic subjects (52 males and 72 females) with culture positive UTI were studied. Patients with negative urine culture (n=64), those diagnosed and treated outside (n=83) and not willing to participate in the study (n=24) were excluded. Results Almost 30% of the patients (both diabetics and non-diabetics) presented with asymptomatic bacteriuria and the prevalence of pyelonephritis was significantly higher (p-0.04) in diabetics compared to non-diabetic patients. Majority of the diabetics with UTI (87.14 %) had HbA1c > 6.5 % with p < 0.001. The isolation rate of Escherichia coli from urine culture was higher (64.60%) among diabetic patients followed by Klebsiella (12.10%) and Enterococcus (9.90%). The prevalence of extended-spectrum beta-lactamase (ESBL) producing E.coli was significantly higher in diabetics (p- value 0.001) compared to non-diabetics. E.coli showed maximum sensitivity to carbapenems in both diabetic and non-diabetic subjects and least susceptibility to ampicillin. Conclusion The prevalence of pyelonephritis is significantly higher in diabetics than in non-diabetic subjects, with E. coli being the most common isolate. Elevated glycosylated hemoglobin predisposes diabetics to UTI. Investigation of bacteriuria in diabetic patients for urinary tract infection is important for treatment and prevention of renal complications.