To describe the effectiveness of the Newborn Life Support (NLS) course in terms of attendees' knowledge, perceived self-efficacy, and clinical applicability. We conducted an electronic survey of NLS course attendees (NLS + group). A control group (NLS-) was recruited via our regional perinatal network. The survey data were analyzed anonymously. Multiple linear regression analysis examined the following: NLS course, job tenure, maternity level, and profession. The survey completion rate was 62% (200/323) for the NLS + group. Among participants, 84% had participated in neonatal resuscitation since their course. The scores for positive perceived experience for neonatal resuscitation (fluency, security, and quality of care delivered) were higher in the NLS + group than the NLS- group (p<.006). After adjustment, the independent factors associated with a higher positive perceived experience were the NLS course, work in tertiary level maternity ward, and job tenure >5 years. The multiple-choice questions (MCQs) score (n = 10) was 8.2 ± 1.3 (NLS+) vs. 6.7 ± 1.5 (NLS-) (p<.0001). NLS course, medical degree, and work in a tertiary level maternity ward were independently associated with higher knowledge scores. The NLS course was associated with a positive perceived experience regarding neonatal resuscitation.Key notesNeonatal resuscitation training programs, like the Newborn Life Support (NLS), have been developed to improve the management and outcomes of newborns with poor adaptation at birth.The NLS course was associated with better knowledge of, and a positive perceived experience (fluency, safety, and quality of care delivered) regarding, neonatal resuscitation.Participation to the NLS course seems to strengthen the perceived self-efficacy in healthcare professionals, which is critical to performing neonatal resuscitation.