Abstract: Background: Central venous catheters (CVCs) play an integral role in the management of many critically ill children. Purpose: The purpose of the study was to assess the effect of nursing care bundle on central venous line associated blood stream infection. Design: experimental design was used. Setting: It was conducted at the pediatric intensive care unit (PICU) in Menoufia University Hospital. Sample: Purposive sample of 33nurses and 34 children with central venous lines. Instruments: Three data collection instruments were used: structured interviewing questionnaire of nurses' knowledge about central line and care bundle, Observational checklist for nurses' practice and Laboratory investigation record. Results: the result of this study showed that there were very highly statistical significant differences between nurses' knowledge on posttest compared to pretest 25.3333±1.81430Vs18.6061±4.14532 respectively as well as nurses' practices on posttest compared to pretest were25.30±1.99Vs22.20 ±1.81 respectively. Also, children who received bundle of care had decreased the rate of blood stream associated infection than children who gotroutinehospitalcare13.17±0.65Vs10.50±0.43respectively. Conclusion: It was concluded that implementation of care bundle of central venous line improved nurses' knowledge and practices regarding prevention of central venous line associated bloodstream infections on post and follow-up tests than pretest. Also, it contributed to lower levels of bloodstream associated infection in children on posttest than pretest. Recommendation: ongoing in-service education programs about nursing care bundle for central venous line should be developed and provided for pediatric nurses working in pediatric intensive care units.