Urinary tract infections (UTIs) are among the most prevalent ailments in the Indian community. The susceptibility and spread of UTI-causing pathogens vary with time and location. This study aims to determine the frequency of UTI-causing microorganisms and the patterns of antibiotic sensitivity in patients from the Telangana region. This observational, prospective study was conducted over six months at a tertiary care hospital in Hyderabad, Telangana. A total of 105 clean-catch midstream urine samples were collected, and each specimen underwent Gram staining for microscopy. The isolates were then evaluated for the presence of urinary tract infections using standard culture techniques, biochemical reactions, and antibiotic susceptibility tests according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Extended-spectrum β-lactamase (ESBL), metallo-β-lactamase (MBL), and AmpC β-lactamase production were detected using double-disc synergy tests (DDST), imipenem-imipenem/EDTA disc tests, and disc-diffusion tests, respectively. Of the samples, 92.5% were identified as Gram-negative bacteria, while 7.5% were Gram-positive. The study observed a predominance of female patients (76.2%) compared to males (23.8%). Among the culture-positive samples, 62.5% were Escherichia coli, 22.5% were Klebsiella pneumoniae, and 7.5% were Proteus mirabilis. The prevalence of ESBL, AmpC, and MBL production was 20%, 7.5%, and 2.5%, respectively. In ESBL screening, E. coli showed the highest incidence (28%), followed by K. pneumoniae (11.1%). The antibiogram of E. coli, the predominant isolate, revealed the highest sensitivity to Amikacin (96%), Gentamicin (88%), Cefoxitin (88%), and Piperacillin-Tazobactam (84%), with lower sensitivity to Amoxicillin-Clavulanic Acid (56%) and Ampicillin (48%). The findings underscore a high prevalence of E. coli and K. pneumoniae among culture-positive samples. The study highlights the importance of identifying multidrug-resistant strains for effective therapy in bacterial infections, thereby reducing the risk of multidrug resistance in hospital and community settings.