Introduction: Blood collection is crucial for disease diagnosis and treatment in hospitalized children. Compared with adults, pediatric blood collection is more challenging due to poor vascular conditions, psychological factors, and family anxiety . Whether blood collection through peripheral venous catheters is feasible remains controversial. Objectives: To retrospectively analyze the impact of different peripheral venous blood collection methods on results and provide evidence for selecting appropriate collection methods in pediatric patients. Methods: We retrospectively analyzed 3,350 pediatric inpatients who underwent peripheral venous blood collection at a tertiary children's hospital from January to December 2024. Basic information was collected through the nursing information management system, and blood specimen results were obtained via Limsbox laboratory management software. Among them, 1,612 cases used direct blood collector collection (1,570 qualified, 42 unqualified), and 1,738 cases used first-time incidental collection through peripheral venous catheters (1,702 qualified, 36 unqualified). Results: The total qualification rate was 97 .4% in the blood collector group (unqualified rate 2.6%) and 97 .9% in the catheter group (unqualified rate 2.1%). Statistical analysis showed no significant difference between the two methods in key quality indicators including anticoagulant specimen coagulation and specimen hemolysis (P > 0.05). Conclusions: First-time incidental blood collection through peripheral venous catheters does not affect the qualification rate of blood collection results. It reduces pain from repeated punctures, improves nursing efficiency , and enhances satisfaction among children and their families.