Urinary tract infections are the most common bacterial infections in humans, both as community-acquired and healthcare-associated infections. The present study was aimed to determine the distribution and antimicrobial susceptibility of strains isolated from urinary tract infections. A hospital cross-sectional based study includes 150 clinically suspected cases of urinary tract infections attended different hospital in Khartoum. Midstream urine samples were collected and pure isolates of pathogenic microorganisms were characterized by colony morphology, gram-stain and standard biochemical procedures. A standard method of agar disc diffusion susceptibility testing method was used to determine susceptibility patterns of the isolates. The most common pathogens isolated were E. coli (57.3%), K. pneumonia (16.7%), P. aeruginosa (14%), E. feacalis (4%). C. albicans (8%). For all UTI isolates, least resistance was observed, E. coli and Klebsiella pneumoniae showed the highest percentage of resistance to ampicillin, Cefepime, Amikacin and Ciprofloxacin, followed by Gentamycin and Imipenem. Significant bacteriuria has been observed. Majority of the isolates were resistant to the commonly prescribed antibiotics. Identifying the pathogen, choosing correct antibiotics, limiting their excess use, improving resistance surveillance systems will help controlling this global problem.