〔Abstract〕 Objective To analyze the distribution of urinary bacteria and drug sensitivity test results in patients with urethral stone obstruction combined with hydronephrosis, and provide a certain reference basis for clinical treatment. Methods A total of 70 patients with urethral stone obstruction and hydronephrosis admitted to the department of urology at Chashan Hospital of Dongguan from May 2021 to August 2023 were selected as the study subjects. All patients underwent urine collection and pathogenic bacteria were cultured in a sterile environment, followed by drug sensitivity tests. The distribution of bacteria and drug sensitivity test results were recorded. Results A total of 70 pathogenic bacteria were isolated from patients with urethral stone obstruction and hydronephrosis, including 50 Gram negative bacteria (71.43%), 16 Gram positive bacteria (22.86%), and 4 other bacteria (5.71%). The majority of detected pathogens were Escherichia coli in 30 (42.86%), Proteus mirabilis in 9 (12.86%), Klebsiella pneumoniae in 6 (8.57%), Enterococcus faecalis in 4 (5.71%), and Streptococcus agalactiae in 4 (5.71%). Escherichia coli has a resistance rate of 100.00% to imipenem, ertapenem, and tigecycline, while Proteus mirabilis and Klebsiella pneumoniae subspecies pneumonia have a resistance rate of over 50.00% to most antibiotics. Enterococcus faecalis has a resistance rate of 100.00% to erythromycin, and Streptococcus agalactiae has a resistance rate of over 50.00% to most antibiotics. Conclusion There are many types of pathogenic bacteria in patients with urethral stone obstruction and hydronephrosis, and there are significant differences in the resistance of different pathogenic bacteria to various antibiotics. During clinical treatment, attention should be paid to evaluating the susceptibility of patients to pathogens and selecting appropriate antibiotics based on the level of drug resistance.