BackgroundCentral venous catheter-related bloodstream infections (CLABSIs) are a significant concern in intensive care units (ICUs) as they lead to increased morbidity, mortality, and healthcare costs. Fortunately, these infections are largely preventable through strict adherence to CLABSI prevention guidelines. Nurses play a critical role in preventing CLABSIs.AimThis study aimed to investigate factors affecting critical care nurses' knowledge, attitudes, and perceived barriers related to implementing CLABSI prevention guidelines, and to predict factors influencing compliance with these guidelines.MethodsThis cross-sectional study was conducted from April to May 30, 2023, with a convenience sample of 470 critical care nurses from ICUs across eight hospitals in Sana’a, Yemen. Data were collected using an observational checklist and self-administered questionnaire. Descriptive statistics, Independent Student’s t-test, one-way ANOVA, Pearson’s correlation coefficient, multiple linear regression, and multilayer perceptron neural networks were performed.ResultsCritical care nurses exhibited low knowledge of CLABSI prevention guidelines, with compliance reaching an acceptable level. Despite the higher perceived barriers, the nurses demonstrated a positive attitude. Nurses with greater knowledge and positive attitudes displayed higher compliance levels. However, perceived barriers were negatively associated with knowledge and compliance. Notably, multilayer neural network analysis identified knowledge and perceived barriers as the strongest predictors of nurses' compliance.ConclusionThe current findings emphasize the need for multifaceted strategies to implement the CLABSI prevention guidelines. These strategies should address knowledge gaps, support positive attitudes, and address practical barriers faced by nurses to ensure successful implementation of CLABSI prevention.