Background: Hand hygiene with ABHR is more effective, time-saving and skin-friendly than traditional handwashing. However, alcohol-based hand rub (ABHR) use can be an obstacle for specific religions. The study aimed to test the effect of a modified hand hygiene program featuring religion-relevant, culturally-specific interventions on compliance and beliefs about ABHR use. Methods: A cohort study design was employed between August and November 2017 at a large tertiary hospital in the United Arab Emirates. Hand hygiene audit data and a cross-sectional survey were used to measure the impact of the modified hand hygiene program on compliance with, and beliefs about, ABHR use. Results: The intervention group had higher mean compliance with overt (p = 0.002) and covert (p = 0.04) ABHR use post-intervention, and higher mean compliance with overt ABHR use (p = 0.002) than the control group. The intervention group had more positive religious beliefs (p = 0.01) about ABHR use than the control group. Conclusion: Implementing a hand hygiene program featuring tailored religion-relevant interventions in a culturally appropriate way had a positive effect on compliance with, and beliefs about, ABHR use.