The aim of the study was to characterize the microbial landscape in the urine of children treated in multi-discipline hospitals based on the spectrum and drug-resistance of the urinary tract bacteria. Materials and Methods. The bacterial strains isolated from the urine of patients aged 1 month to 18 years with urinary tract infections treated in the pediatric wards of Nizhny Novgorod clinics from 2005 to 2017 were studied. The degree of bacteriuria and sensitivity to antibiotics/antibacterial agents were determined. Results. The results indicated a change in the spectrum of pathogens in urine samples of patients that underwent treatment in the Children's City Clinical Hospital No.1 (Nizhny Novgorod) during 2000-2016. A mixture of microbial species that prevailed earlier has been replaced with bacteria monocultures in recent years (2014-2016). Among those, gram-negative bacteria dominated (E. coli, Klebsiella, Proteus); the gram-positive flora was represented by the Streptococcus, Enterococcus, and Staphylococcus genera. Since 2014, the occurrence rate of Klebsiella has increased to 14.7%. In the surgery department of the Regional Children's Clinical Hospital of Nizhny Novgorod, gram-negative bacteria monocultures dominated throughout the entire study period (2005-2017). Among the gram-positive flora, bacteria of the Staphylococcus genus took the lead. In recent years, the occurrence of Klebsiella has also increased. Conclusion. Over the past 16 years, a change in the spectrum of the urinary tract pathogens occurred in both pediatric and surgery departments of multi-discipline children's hospitals. Continuous monitoring of urinary microflora and its sensitivity to antibiotics and other antimicrobial drugs is important for adjusting urinary tract infections therapy in pediatric patients.