Background: Urinary tract infections (UTI’s) define a condition in which urinary tract is infected with a pathogen causing inflammation. Urinary tract infections are one of the most prevalent infections affecting people of all age group from neonates to adults and are major cause of morbidity and several disorders in patients. In serious cases urinary tract infection lead to kidney failure, septicemia, bacterial endocarditis, prostatitis and infertility. Aim: The objective of this work was to the study the prevalence of urinary tract infections among patients of Ujjain and to identify prominent bacterial pathogens responsible for causing UTI. Methods: A total of 500 urine samples were collected aseptically and cultured on Blood agar and Mac’conkey agar. The bacterial isolates were identified based on their colony morphological characteristics, Gram stain reaction and their biochemical tests. Result: After overnight incubation a total of 307 out of 500 patients tested positive for UTI suggesting that (61%) of patients suffered from UTI infections. It was seen that (89%) of bacterial isolates causing UTI were Gram negative and (11%) were Gram positive. The identification of pathogens indicated that E. coli is most prevalent uropathogen followed by K. pneumonae, P. aeruginosa, E. faecalis, S. aureus and P.vulgaris. Conclusion: The study shows that prevalence of UTI is higher in females as compared to males. It was also been seen that UTI infections were prevalent in urban as well as in rural patients, however, the incidence of infection was slightly high in urban patients. This study is important as constant survey and identification of uropathogens is essential for effective treatment of UTI.