There are an estimated 150 million cases of urinary tract infection (UTIs) reported each year, putting it among the most prevalent infectious diseases. The study aimed to estimate the prevalence of AmpC β-lactamase and Extended spectrum β-lactamase (ESBL) production amidst E. coli and Klebsiella pneumonia strains from UTI cases. A total of 406 non-repetitive urine samples (203 isolates of E. coli and K. pneumoniae each) were analyzed. The study revealed significant prevalence of MDR strains, with 157 isolates (77%) of E. coli and 171 isolates (84%) of Klebsiella showing resistance to multiple antibiotics. 195 isolates (96%) of E. coli and 179 isolates (88%) of K. pneumoniae showed highest levels of resistance to Ciprofloxacin and Cefoxitin, respectively. Production of ESBL was found in 128 isolates (63%) of E. coli and 75 isolates (37%) of K. pneumoniae by Disc diffusion method, while Putative AmpC was seen in 103 isolates (51%) of E. coli and 120 isolates (59%) of K. pneumoniae by Combination disc test. These findings highlight the immediate need for robust antibiotic stewardship practices and effective strategies for infection control to manage UTIs, particularly against MDR strains, adopting targeted antibiotic treatment based on patient-specific factors and local resistance pattern.