Spina bifida is a birth condition that results in an improperly formed portion of the spinal column, which exposes a portion the spinal nerves and spinal cord through a hole at the back. The nurse is crucial in immediate assessment of neonates after birth, positioning, and lesion stabilization. The study aimed to evaluate the effect of nursing intervention on the performance of nurses regarding care for newborns with spina bifida. Design: a quasiexperimental design was used. Setting: Neonatal intensive care unit at Tanta University Main Hospital. Subject: a convenience sampling of 70 nurses participated in the study. Two tools were utilized: Tool I Structured interview schedule that comprised of two parts and Tool II: Nurses, performance Observational checklist related to preoperative and postoperative care for neonates with spina bifida. The results revealed that the study group's means of total knowledge and mean of total practice regarding the pre- and postoperative care of newborns with spina bifida improved immediately after and one month after the execution of the educational program. The study concluded that the implementation of the program resulted in significant improvements in nurses' knowledge and practice, so that the program successfully corrected the nurses' lack of knowledge and performance. The study recommended that holding in-service training sessions regularly to maintain nurses' skills and knowledge current.