Spiritual care is essential to the well-being of patients, and nurses provide spiritual care as a fundamental part of nursing practice. In this study, we investigated the spiritual care needs of hospitalized patients to determine whether the perceived knowledge of nurses corresponded with these spiritual care needs. A cross-sectional study was conducted on 1351 hospitalized patients and 200 registered nurses recruited from a medical center in central Taiwan. A questionnaire, including the 21–item Spiritual Care Needs Inventory (patient and nurse version) and basic demographic information, was distributed to eligible participants. The top three items of the spiritual care needs expressed by the hospitalized patients were respect for privacy and dignity, showing concern, and guidance in gaining a sense of hope in life; the percentages of nurses not knowing how to provide these spiritual care needs were 0%, 1%, and 15%, respectively. The spiritual care needs of patients showed a significant relationship with the knowledge of nurses, suggesting that the perceived knowledge of the nurses generally corresponded with the spiritual care items that the patients required most.