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Neonatal intensive care units (NICUs) are caring for an increasing number of infants born with neonatal abstinence syndrome (NAS). The literature identifies the need for education for NICU nurses on NAS including skills for interacting with the mother with substance use disorder.An evidence-based practice project was developed to offer an educational presentation targeting these topics to 206 NICU participants (93% registered nurses, 1% licensed vocational nurses, and 6% nursing assistants) at a level IV NICU.A pretest/posttest was developed to assess knowledge of the participants prior to and after completion of the educational presentation. A posteducational questionnaire was developed to evaluate the skills learned for interacting with the mother with substance use disorder and the participant's professional readiness.The posttest demonstrated a statistically significant (P < .001) increase in knowledge of NAS. On the post-educational questionnaire, 96% of participants correctly identified 3 skills they would use clinically with mothers with substance use disorder and 84% of participants identified 2 personal strengths and 2 weaknesses that influence their care of infants with NAS and their families (professional readiness).These results demonstrate that this educational presentation was effective and should be replicated at other facilities to improve the knowledge and skills of NICU nurses to promote improved care for infants with NAS.Future studies should examine the impact on patient outcomes by preforming post-discharge interviews with the mothers of infants with NAS in the NICU pre- and post-educational intervention. Expanding the presentation into an interprofessional educational opportunity would promote increased knowledge and care by the entire multidisciplinary team.