Introduction: Peripherally inserted central catheters (PICC) are widely used in neonatal intensive care units (NICUs) for longterm intravenous therapy , but catheter tip malposition is a common complication that can lead to severe outcomes such as phlebitis, thrombosis, pleural effusion, and even pericardial tamponade. Although evidence-based guidelines for preventing and managing PICC tip malposition exist, clinical implementation remains limited due to poor evidence translation, low adherence, and inconsistent management processes. Objective: To develop an evidence-based practice protocol for the prevention and management of PICC tip malposition in neonates and evaluate its effectiveness. Methods: Guided by the Knowledge-Action Framework, this study screened and selected 29 best pieces of evidence through a Delphi expert consultation process. Baseline audits were conducted on 38 neonates and 29 nursing staff from May to September 2024. Knowledge and practice gaps were identified using a questionnaire and observational checklist. Barriers and facilitators were assessed based on the Consolidated Framework for Implementation Research (CFIR). An evidence-based practice protocol was developed considering these factors, followed by clinical implementation and evaluation between October 2024 and February 2025. Results: Following the intervention, nurses' knowledge scores improved from 58.00±14.55 to 78.45±11.18 (P < 0.05), and the compliance rate with audit criteria increased significantly (P < 0.05). The incidence of PICC tip malposition decreased significantly (P < 0.05). Conclusion: The evidence-based practice protocol significantly improved nursing knowledge, standardization of care, and reduced the incidence of PICC tip malposition in neonates, providing a practical model for clinical practice in NICUs.