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Background Effective provider-patient communication is crucial to the delivery of high-quality care. Communication roadblock such as righting reflex is widely observed among providers and can lead to relational disengagement. In previous work, nurses felt ill-equipped to communicate effectively with HIV-positive patients to support medication adherence. Providing nurses with continuing education opportunities to improve their relational skills is a major target for optimizing the quality of care. Virtual patient simulation is one promising strategy that needs to be evaluated among graduate nurses. This study aimed to assess the acceptability of a virtual patient simulation to improve nurses’ relational skills in a continuing education context. Methods We conducted a convergent mixed methods study by combining a quantitative pre-experimental, one-group post-test design and a qualitative exploratory study. We used convenience and snowball sampling approaches to select registered nurses (n = 49) working in Quebec, Canada. Participants completed an online sociodemographic questionnaire, consulted the automated virtual patient simulation (informed by motivational interviewing), and filled out an online post-test survey. Descriptive statistics (mean, SD, median, interquartile range) were used to present quantitative findings. From the 27 participants who completed the simulation and post-test survey, five participated in a focus group to explore their learning experience. The discussion transcript was subjected to thematic analysis. At the final stage of the study, we used a comparison strategy for the purpose of integrating the quantitative and qualitative results. Results Nurses perceived the simulation to be highly acceptable. They rated the global system quality and the technology acceptance with high scores. They reported having enjoyed the simulation and recommended other providers use it. Four qualitative themes were identified: motivations to engage in the simulation-based research; learning in a realistic, immersive, and non-judgmental environment; perceived utility of the simulation; and perceived difficulty in engaging in the simulation-based research. Conclusions The simulation contributed to knowledge and skills development on motivational interviewing and enhanced nurses’ self-confidence in applying relational skills. Simulation holds the potential to change practice, as nurses become more self-reflective and aware of the impact of their relational skills on patient care. Trial registration ISRCTN18243005, retrospectively registered on July 3 2020.