Objective: To investigate the distribution, drug resistance of pathogens and prognostic risk factors in patients with positive catheter culture, and to provide pathogenic basis and preventive measures for clinical catheter-related infection. Methods: The etiological data of patients with positive catheter culture in the First Affiliated Hospital of Nanjing Medical University from 2018 to 2023 were retrospectively analyzed, and the clinical data of the patients with the top three pathogens were collected to analyze the risk factors affecting the prognosis. Results: The overall positive rate of catheter culture was 14.7% (1240/8407). After removing the same pathogens isolated from the same patient, a total of 1096 strains of pathogens were isolated from 916 patients, 48.7% of which were gram-negative bacteria, mainly Klebsiella pneumoniae, Acinetobacter baumannii and Escherichia coli. Gram-positive bacteria accounted for 39.2%, mainly Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus. The resistance rate of Klebsiella pneumoniae to most antibiotics was higher than that of Escherichia coli. The resistance rate of Acinetobacter baumannii to most cephalosporins and carbapenems was higher (>85.0%). The detection rates of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis were 35.6% and 92.2%, respectively. No vancomycin and linezolid resistant Staphylococcus and Enterococcus were detected. Multivariate analysis showed that male, ≥65 years old, history of dialysis, combined catheter-associated bloodstream infection and tracheal intubation were independent risk factors for prognosis in patients with catheter-associated infection (P<0.05). Conclusion: The incidence of catheter-associated infection is high, which needs clinical attention. Continuous monitoring and analysis of etiological characteristics in the local area can provide theoretical basis for clinical empirical anti-infection treatment.