Background: Central venous line infection (CVLI) are linked to higher rates of sickness, death, and medical expenses and represent a major risk to patient safety. Nurses play a crucial part in preventing CVLI, Through commitment to evidence-based practice. Aim of the study: This study aimed to assess the nurses’ performance regarding prevention of central venous line infection. Subjects and methods: Research design: Descriptive research design. Setting: The study was conducted in surgical intensive care units present at New Surgery Hospital at Zagazig University Hospitals. Subjects: A convenient sample of all nurses that are working in the above-mentioned setting (75 nurses). Tools of data collection: Two tools were used, Self-administered questionnaire and an observation checklist. Results: This study revealed that 54.7% and 69.3% of the studied nurses had unsatisfactory overall knowledge and practice levels regarding prevention of CVLI, respectively. Conclusion: Over half of studied nurses had unsatisfactory overall knowledge about prevention of CVLI and over two-thirds had unsatisfactory overall practice levels. Furthermore, a significant relation was found between the nurses' overall knowledge and practice. Recommendations: Effective and standardized training program that are based on effective interventions can enhance nurses' knowledge and practice application in a variety of healthcare settings and need to be updated on a regular basis. Keywords: Central venous line, Infection, Nurses’ performance, Prevention.