The aim of this study was to explore the prevalence and antimicrobial susceptibility pattern of UTI among hospitalized children in our center. Records were reviewed retrospectively to discern the urine culture tests and antibiotic susceptibility tests of children with UTI from January 2014 to December 2021. Consequently, a total of 840 hospitalized children with UTI were collected. The proportion of culture-positive UTI was 54.52% (458/840) with majority of the isolates being Gram-negative bacteria. There were 694 children with complicated UTI in those children. The proportion of culture-positive UTI was 397/694 (57.20%). Gram-negative bacteria mainly included Escherichia coli 121 (30.48%), Klebsiella pneumoniae 32(8.06%), Pseudomonas aeruginosa 20 (5.04%), Enterobacter cloacae 11 (2.77%) and Citrobacter 13 (3.28%). Escherichia coli isolates were highly sensitive to amikacin (95.87%), ertapenem (91.74%), nitrofurantoin (92.56%), imipenem (90.08%), and cefotetan (87.60%) and high rate of resistant were also detected to ampicillin (93.39%), cephazolin (76.03%), and ceftriaxone (73.55%). Klebsiella pneumoniae isolates showed sensitive to ertapenem (96.87%), amikacin (96.87%), imipenem (93.75%) and piperacillin-tazobactam (90.62%), while highly resistant were observed to ampicillin (96.87%), cephazolin (75.00%), and ceftazidine, ceftriaxone, aztreonam (each 62.50%). The isolated Gram-positive bacteria mainly contained Enterococcus faecalis (14.61%) and Enterococcus faecium (each 16.62%). Enterococcus faecalis were sensitive to vancomycin, penicillin-G, tigecycline, linezolid and nitrofurantoin (100%, 94.83%, 88.48%, 87.93%, 86.20, respectively) and resident to tetracycline (86.21%), quinupristin (84.48%), erythromycin (75.86%). Likewise, Enterococcus faecium were sensitive to vancomycin, linezolid, tigecycline, quinupristin (98.48%, 95.45%, 84.85% , 83.33%), while resident to ampicillin (93.94%), erythromycin (89.39%), penicillin-G (84.85%), and ciprofloxacin (77.27%). In addition, there were 146 children diagnosed simple UTI. The culture-positive UTI was 61/146 (41.78%). The predominant bacteria were Escherichia coli (73.77%, 45/61) followed by Enterococcus faecium and CoNS (each 6.56%). Escherichia coli were highly sensitive to ertapenem, imipenem, and cefotetan (each 100%), amikacin (95.56%) and nitrofurantoin (91.11%) and high rate of resistant were also detected to ampicillin (91.11%), and trimethoprimsulfamethoxazole (64.44%). Moreover, both UTI organisms showed a high multiple drug resistance frequency (80.98% and 77.05%). Culture and susceptibility test is vital for appropriate management of UTI in the study. Further large-scale studies are warranted to examine the generalizability of our findings.