Objective To summarize the characteristics and drug resistance changes of urinary tract infection through the analysis of pathogen distribution and drug sensitivity. Methods Pathogens isolated from urine culture samples collected from the People's Hospital of Wuhan University during three years from January 2020 to December 2022 were identified by Bruker mass spectrometer and Phoenix 100 for drug sensitivity analysis, and antimicrobial susceptibility analysis was performed by WHONET 5.6 software. Results The main pathogens of urinary tract infection were Gram-negative bacteria (64.2%), Escherichia coli (39.0%), Klebsiella pneumoniae (10.0%) and Pseudomonas aeruginosa (3.8%); Gram positive bacteria accounted for 28.5%, mainly Enterococcus faecalis (12.7%) and Enterococcus faecalis (9.1%); Gram negative bacteria were more sensitive to colistin, imipenem, meropenem, tigecycline, amikacin and piperacillin / tazobactam; The sensitivity rate of Gram-positive bacteria to glycopeptide antibiotics such as vancomycin，linezolid and teicoplanin is generally more than 90%. Conclusion The drug resistance of bacteria to antibiotics is becoming more and more serious. Therefore, timely summarizing and analyzing the distribution and drug resistance of pathogens of urinary tract infection is of great significance for controlling the spread of drug-resistant strains and guiding clinical rational drug use