OBJECTIVES:  Urinary tract infections (UTIs) often remain one of the most common infections diagnosed in outpatients. The purpose of the present study is to determine the prevalence of UTI, isolation and identification of UTI causing bacteria from urine samples to common used antibiotics in a tertiary care hospital, Abbottabad, Pakistan. METHODS:  Nine hundred and seventy one patients with clinical symptoms of UTIs were examined. The disc-diffusion method was used to carry out test for antibiotic resistance of organisms. Commercially available discs of antibiotics were used, each having varying mode of action like inhibitors of protein synthesis, DNA synthesis inhibitor and cell wall synthesis inhibitors. RESULTS:  Gram-negative microorganisms among total isolated organisms were 21%, whereas 15% were found to be Gram-positive. The most prevalent Gram-negative organisms were Escherichia coli (20%) followed by Klebsiella pneumonia (1%) while among Gram-positive microorganisms, Staphylococcus aureus (10%) and Staphylococcus saprophyticus (3%) were the most prevalent organisms followed by streptococci (2%)The most effective antibiotics for treatment of Gram-negative isolates were Imipenem (74.74%), Piperacillin-sulbactam (75%), in addition to Cefotaxime, Ceftriaxone, and Meropenem that showed 100% sensitivity. Among Gram-negative microorganisms, Amoxicillin was highly efficacious (89%) followed by doxycycline (78%). Piperacillin-sulbactam, Cefotaxime and sulfamethoxazole/trimethoprim showed 100% sensitivity against Candida albicans