Central venous catheters (CVCs) play an integral role in the management of many critically ill children. Aim: To assess the effect of nursing care bundle on central venous line-associated bloodstream infection. Design: a quasi-experimental design. setting: It was conducted at the pediatric intensive care unit (PICU) in Menoufia University Hospital. The sample included 33 nurses and 34 children with central venous lines. Data collection instruments: structured interviewing questionnaire, Observational checklist for nurses' practice, and Laboratory investigation record. Results: A very highly statistically significant differences between nurses' knowledge on post-test compared to pretest) as well as nurses' practices on post-test compared to pretest. Conclusion: Implementation of care bundle of central venous line improved nurses' performance regarding prevention of central venous line-associated bloodstream infections on the post and follow-up tests than pretest. Recommendations: Ongoing in-service education programs about nursing care bundles for central venous lines should be developed and provided for pediatric nurses working in pediatric intensive care units.