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Abstract Background: Surfactant Replacement therapy plays a crucial role in reducing morbidity and mortality associated with respiratory distress syndrome among preterm infants. Aim: To assess nurses’ knowledge and skills regarding surfactant administration for preterm infants. Methods: A descriptive research design was conducted at two neonatal intensive care units at El Manial University Hospital (Kasr Al-Aini) and Cairo University Children’s Hospital (El Monira) from December 2024 to May 2025. A convenient sample of fifty bedside nurses working in the previously mentioned settings was included. Data were collected using two tools: a structured questionnaire to assess nurses’ knowledge and an observational checklist to evaluate nurses’ skills related to surfactant administration. Results: The study findings revealed that only a small proportion of nurses achieved an overall satisfactory level of knowledge regarding respiratory distress syndrome (RDS) and surfactant administration. Although many nurses demonstrated satisfactory knowledge related to RDS, knowledge concerning surfactant administration remained unsatisfactory. Furthermore, none of the studied nurses demonstrated a satisfactory level of skills related to surfactant administration .A moderate positive correlation was found between nurses’ knowledge and skills scores. Educational qualification demonstrated a strong positive association with both knowledge and skills levels. Conclusion: The study concluded that the level of nurses' knowledge and skills regarding surfactant administration for preterm infants was unsatisfactory. A positive association was found between nurses’ knowledge and their clinical skills. Recommendation: Regular assessment programs and continuous educational training should be implemented for nurses working in neonatal intensive care units to enhance their knowledge and clinical skills related to surfactant administration.