Neonatal resuscitation is a life-saving intervention for birth asphyxia, a leading cause of neonatal mortality. Worldwide, four million neonate deaths happen annually, and birth asphyxia accounts for one million deaths. Improving providers' neonatal resuscitation skills is critical for delivering quality care and for morbidity and mortality reduction. However, retention of these skills has been challenging in developing countries, including Ethiopia. Hence, this study aimed to assess neonatal resuscitation skills retention and associated factors among midwives and nurses in Eastern Ethiopia. An institution-based cross-sectional study was conducted using a pre-tested, structured, observational checklist. A total of 427 midwives and nurses were included from 28 public health facilities by cluster sampling and simple random sampling methods. Data were collected on facility type, availability of essential resuscitation equipment, socio-demographic characteristics of participants, current working unit, years of professional experience, whether a nurse or midwife received refresher training, and skills and knowledge related to neonatal resuscitation. Binary logistic regression was used to analyse the association between neonatal resuscitation skill retention and independent variables. About 11.2% of nurses and midwives were found to have retention of neonatal resuscitation skills. Being a midwife (AOR, 7.39 [95% CI: 2.25, 24.24]), ever performing neonatal resuscitation (AOR, 3.33 [95% CI: 1.09, 10.15]), bachelor sciences degree or above (AOR, 4.21 [95% CI: 1.60, 11.00]), and good knowledge of neonatal resuscitation (AOR, 3.31 [95% CI: 1.41, 7.73]) were significantly associated with skill retention of midwives and nurses. Basic neonatal resuscitation skills of midwives and nurses in Eastern Ethiopia are not well retained. This could increase the death of neonates due to asphyxia. Being a midwife, Bachelor Sciences degree or above educational status, ever performing neonatal resuscitation, and good knowledge were associated with skill retention. Providers should be encouraged to upgrade their educational level to build their skill retention and expose themselves to NR. Further, understanding factors affecting how midwives and nurses gain and retain skills using high-level methodology are essential.