Objective: This study aimed to determine the knowledge and compliance among paediatric nurses regarding the prevention of central line-associated bloodstream infections. Methods: The research employed a descriptive correlational approach in the government-subsidised hospitals of the Hail Region of Saudi Arabia. The participants were the staff nurses responsible for the insertion of the central line.Results: Results showed that there were significant differences between gender with knowledge (t =-2.701; p = 0.007) and compliance (t =-3.118; P = .002), civil status with knowledge (t = 7.03; P <0.000) and compliance (t = 2.450; P = 0.015), educational background with knowledge (t = 10.938; P <0.000) and compliance (t = 11.890; P <0.000), age with knowledge (F = 63.376; P <0.000) and compliance (F = 21.178; P <0.000), and the years of working experience with knowledge (F = 101.390; P <0.000) and compliance (F = 82.452; P <0.000). It is noteworthy that there was a correlation between the knowledge and compliance of nurses on central line-associated bloodstream infections (CLABSI) (r = .552; P <0.000).Conclusion: Gender, civil status, educational background, age and working experience were significant to knowledge and compliance. There was also a significant correlation between the knowledge and compliance of nurses on CLABSI. Therefore, programs that aim to increase nurses' knowledge and compliance with CLABSI preventive protocols need to take a number of criteria, including the nurses' gender, civil status, educational background, age and amount of previous professional experience into consideration. Given the wide range in both the CLABSI and the compliance rates across the participating hospitals, there is likely still opportunity for improvement in nurses' knowledge and compliance.