Background: The most important psychological problems of dialysis patients are anxiety, depression, and stress. These psychological problems cause different consequences for the patients. Therefore, it seems necessary to use strategies to reduce the psychological problems in hemodialysis patients. Objectives: This study was conducted to evaluate the effectiveness of spiritual therapy on depression, anxiety, and stress in hemodialysis patients referred to the dialysis ward of special diseases center of Birjand in 2019 - 2020. Methods: In this semi-experimental study, 70 patients with chronic kidney disease (CKD) under hemodialysis referred to the dialysis ward of special diseases center of Birjand in 2019 - 2020 were selected by available sampling method and randomized into two experimental groups (35 patients) and control (35 patients). The patients in the experimental group received eight sessions of 60-minute (one session per week) of spiritual intervention therapy, and the control group received no spiritual intervention. Data collection tools were a form of demographic profile and Lovibond SH and Lovibond PF (1995) questionnaire. Data were analyzed using SPSS 26 statistical software, and Fisher’s exact test, chi-square test, independent t-test, repeated measures ANOVA, Bonferroni post hoc test at a significance level of P < 0.05. Results: In this study, 68 patients were examined in two experimental (33 patients) and control (35 patients) groups. The mean score of depression, anxiety, and stress before the intervention in patients in the experimental and control groups was not significantly different (P = 0.61), but immediately and three months after the intervention in patients in the experiment group was significantly lower than the control group (P < 0.001). Conclusions: Based on the results of the present study, it can be said that spiritual therapy is considered an effective solution for reducing depression, anxiety, and stress in hemodialysis patients.