The prevalence, antimicrobial susceptibility pattern, and associated factors of urinary tract infections among Punjabi patients attending Tertiary Care Hospitals of Punjab were the objectives of this study. Participants were selected by systematic random sampling in a cross-sectional study at Tertiary Care Hospitals of Punjab. 988 participants provided clean catch midstream urine samples, which were then cultured and tested for antimicrobial susceptibility. We inoculated n=988 urine samples in which 255 samples were found positive for bacterial growth. Among them 126 were E. coli, 66 Klebsiella pneumoniae, 4 Enterobacter cloacae, and 21 were Pseudomonas spp. The frequency of Uropathogens was observed for Gram negative E. coli 49.02% and Gram positive bacteria were 7.78%. The prevalence of urinary tract infections was 16.4%. Pseudomonas aeruginosa (19, 7.3%), Pseudomonas spp. (2, 0.7%), Enterobacter cloacae (4, 1.5%), and Gram positive bacteria such as Methicillin resistant staphylococcus aureus (3, 1.1%), Methicillin sensitive staphylococcus aureus (15, 5.8%), and Enterococcus faecalis (20, 7.7%) were the most common bacteria. Ampicillin and tetracycline were both resistant to Gram-negative bacilli (96%) and Gram-positive cocci (72%), respectively. In the study area, a culture and susceptibility test is essential for effective urinary tract infection management.