Objective: To describe de epidemiological, clinical, and microbiological features of neonatal urinary tract infection. Methods: A descriptive retrospective study of neonates with urinary tract infection admitted to Clinica Universitaria Bolivariana (Medellin, Colombia) between January 2013 and December 2017. Data about urinary tract infection features were collected from the clinical records of the hospital. For data analysis, qualitative variables were presented as absolute and relative frequencies, and quantitative variables were presented as mean and standard deviation or median and interquartile ranges. Results: Sixty-five patients were included. The most common uropathogens were Escherichia coli (52%) and Enterococcus faecalis (20%). The most frequent clinical features were fever (46%), jaundice (38%) and apneas (15%). Of 21 patients, 28% had C reactive protein raised. Urine dipstick test was positive for leukocyte esterase in 65%. There were 15 nosocomial infections (23%). Renal ultrasound was performed in all patients, of which 25% had congenital anomalies of the kidney and urinary tract. Voiding cystourethrogram was performed in 35 patients, of which 17% had vesicoureteral reflux; 67% of them had a normal renal ultrasound and 83% of them had a non-Escherichia coli bacteria isolation. Two patients had bacteremia and one patient had meningitis. Conclusion: neonatal urinary tract infection is a disease with multiple clinical manifestations, where fever and C-reactive protein weren’t common marker of inflammatory response. In this study, having a normal renal ultrasound doesn’t discard the possibility of having vesicoureteral reflux, and other criteria should be considered to select which patients need studies for vesicoureteral reflux.