Background: Increasing resistance to antimicrobials is a worldwide problem. The aim of our study was to determine the pathogens and antimicrobial susceptibility of bacteria causing urinary tract infection (UTI) in children. Methods: This is a prospective cohort study conducted over a 10-month period with 101 children hospitalised at Kamenge University Hospital for acute UTI. The infections were confirmed by Kass urinalysis criteria, and culture and susceptibility antibiotic tests were performed for isolated microbial agents. Results: Frequency of UTI in the overall population of children hospitalised at Kamange University Hospital was 8.4%. Of the 101 children with UTIs, 87 (86.1%) were under the age of 24 months. Diagnosis of pyelonephritis (82%) was the most common, followed by cystitis (18%). Escherichia coli (82%) was the most frequent pathogen causing UTI. We found E coli and Klebsiella pneumonia to be resistant to aminopenicillins (100%), cotrimoxazole (98.2%, 100%), Augmentin (amoxicillin/ clavulanic acid) (70.5%, 80%), cefotaxime (45.8%, 28.6%), cefuroxime (36.8 to 45.5%, 50%), fluoroquinolones (33.3 to 53.6%, 28.6 to 50%), gentamicin (27.5%, 20%), and nitrofurantoin (9.3%, 50%). Conclusion: E coli is the main causal agent of UTI in childhood with a high resistance to antibiotics. Appropriate antibiotics for empiric therapy should be based on local circulating bacterial strains and resistance profiles.