Background <p>The stress associated with the hospitalization of a newborn exerts considerable psychological pressure on parents. In the neonatal intensive care unit, nurses generally focus primarily on providing physical care. Although this care is vital, it is also equally crucial to consider spiritual needs and provide spiritual care to support families. Providing spiritual care to parents is part of nurses’ care. This requires a proper understanding of spirituality and spiritual care.</p> Methods <p>This was a descriptive-analytical study in which nurses’ perceptions of spirituality and spiritual care were ascertained in a national analytical survey in NICUs in Iran. The research sample consisted of 1643 NICU nurses. The sampling method was multistage random sampling. We employed the Spirituality and Spiritual Care Rating Scale for data collection.</p> Results <p>The mean total spirituality and spiritual care score of the nurses was (67.44 ± 9.38). There was a significant relationship between age (<i>p</i> = 0.04), length of clinical experience (<i>p</i> = 0.04), and attention to spiritual care (<i>p</i> ≤ 0.001). A total of 75.71% were interested in spiritual care, but only 31.77% were practicing it. Interest in implementing spiritual care (<i>p</i> &lt; 0.001), implementing spiritual care in the ward (<i>p</i> = 0.02), and familiarity with the concepts of spirituality and spiritual care (<i>p</i> = 0.045) were the best predictors of the total score for understanding spirituality and spiritual care, respectively.</p> Conclusion <p>Overall, NICU nurses are inclined toward spirituality. They believe in the significance of providing spiritual care. Spiritual care is not associated with a particular religion. Nurses are interested in providing this care by receiving adequate training, focusing on the spiritual needs of parents. This study indicated important results for health policy makers, suggesting the necessity of providing spiritual care training to NICU nurses.</p>