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Heart failure (HF) has been emerging as a general health problem over recent decades. Spiritual care is a type of support service provided to patients suffering from HF. Spiritual care intervention in nursing is a unique aspect of care, which cannot be replaced by psychosocial care. Considering the importance of anxiety for caregivers of patients with HF, the present study aimed to examine the effect of spiritual intervention on anxiety in caregivers of patients with HF in Ilam, Iran. This research is a semi-experimental study, 71 caregivers of patients with HF were randomly assigned to experimental group (n = 34) and control (n = 37) group. Beck Anxiety Inventory, which consisted of 21 items and scored based on a 4-point Likert scale ranging from zero to three, was employed to collect data. The scores range from zero to 63 with the higher scores indicating the higher level of anxiety. The experimental group received spiritual intervention over six 45-minute sessions in a period of 2 weeks (14 days; three times a week; every other day). Data were analyzed using descriptive and inferential statistics run in the SPSS software version 16. The result showed a difference between the level of anxiety in two groups after the intervention (P = 0.001). Anxiety level in the experimental group three weeks after intervention (27.88 ± 7.10) was significant in comparison with before intervention (45.06 ± 5.79) (P = 0.001). According to the results, the spiritual intervention reduced the anxiety level in the caregivers of patients with HF. Nurses are recommended to provide such necessary training to caregivers in order to provide the grounds for reducing their anxiety.