BACKGROUND: Urinary tract infection (UTI) is a collective term used to describe the microbial invasion of any part of the urinary tract, which consists of the kidneys, ureters, bladder, and urethra. It is one of the most common bacterial infections in both sexes with a predilection in females. OBJECTIVES: The objectives of the study were to isolate the bacteria causing UTI and the highest resistance microorganisms with resistance rates to different antibiotics. STUDY DESIGN: This was a cross-sectional study.METHODS: A cross-sectional study with a sample size of 1000 patients both inpatient and outpatient referred to the laboratory of Al-Kindy Teaching Hospital for urine culture and antibiotic susceptibility examination. RESULTS: Of 1000 samples, 318 (31.8%) urine samples were found positive while 682 (68.2%) were negative. Females with bacteriuria held the highest percentage (72.3%) over males (27.7%). Most frequent pathogen was Escherichia coli (40.5%) followed by Klebsiella pneumonia (25.7%) which together accounted for 66.2% of the total uropathogens. The uropathogenic bacterial isolates showed different resistance rates to antimicrobial agents: Cefixime (74%), ticarcillin/clavulanic acid (73.87%), rifampin (68.75%), ceftriaxone (60.31%), cefepime (44.44%), ciprofloxacin (43.65%), aztreonam (33.78%), nitrofurantoin (29.61%), gentamicin (25.64%), and amikacin (7.31%) being the most effective with a susceptibility rate of (89.63%). CONCLUSIONS: E. coli remains to be the most frequent bacterial uropathogen causing urinary infections. Effective drug may be in the clinical study, in microbiology is only that bacteria were most susceptible to amikacin. β-lactams have the highest resistance rates, and the most resistance antibiotic in this study is cefixime.