Purpose: Given the crucial role of professional skepticism in enhancing audit quality, this study aimed to examine the effect of religiosity and spirituality on auditors’ professional skepticism, with an emphasis on gender differences.

Method: This applied research employed a descriptive-correlational design using structural equation modeling (SEM). The statistical population included auditors working in audit firms accredited by the Audit Organization and the Securities and Exchange Organization in 2025. Based on Cochran’s formula, 300 participants were selected through simple random sampling. Data were collected using three validated standard questionnaires (religiosity, spirituality, and professional skepticism). The data were analyzed using AMOS and SPSS software, and statistical techniques such as path analysis and SEM.

Findings: The results indicated that both religiosity and spirituality positively influence auditors’ professional skepticism, with religiosity having a stronger effect. Moreover, these effects were more pronounced among female auditors, such that women with higher levels of religiosity and spirituality demonstrated greater professional skepticism.

Conclusion: The findings reveal that religiosity and spirituality play a positive role in enhancing auditors’ professional skepticism and can serve as effective internal resources for promoting ethical behavior. These results highlight the importance of incorporating spiritual and religious components in auditor training programs. In terms of knowledge contribution, the study enriches the behavioral auditing literature by clarifying the link between psychological factors and professional conduct and offers a basis for educational policy-making in auditing institutions.