Healthcare providers without working experience in isolation wards experience enormous challenges. Traditional ward orientation is constrained by space, time, and even infection risk in particular periods (eg, the coronavirus disease 2019 pandemic). Virtual reality has been used widely, but rarely in wards. This study aimed to explore the experience of utilizing virtual reality for isolation ward training among nurses. In this study, nurses completed virtual reality training via an online platform and were then trained in isolation wards, after which their perceptions were explored by questionnaire and interviews. A total of 1868 participants completed the training. Most participants thought the preservice training was important and believed the virtual reality experience was consistent with the in-person training. Virtual reality was found not only to be convenient and valuable for training but also to have the benefits of occupational protection. However, whereas 50.48% of participants wanted to learn the ward via virtual reality, 87.21% of participants wanted to learn via in-person training before working in the wards. As a substitute for in-person training, virtual reality is a feasible and practical instrument to provide preservice training in particular periods. However, there is room for improvement due to general discomfort and technological problems.