Objective To explore the design and practice effect of virtual simulation experiment teaching based on motivation theory. Methods Two hundreds and seven nursing undergraduates were selected by purposive sampling. Then teaching practice through three steps were carried out，including pre-class online self-study，classroom teaching and after-class consolidation，after which，the teaching effect was investigated. Results The times of students logging in the virtual simulation experiments system was 6.43±2.10，the median duration was 35 min，the first experiment test score was 73.01±28.58，the highest experiment test score was 95.05±3.38，and the total experimental teaching score was 88.64±7.51. The students’ self-assessment of the benefit degree of virtual simulation experiment learning was 4.20，the perceived interest was 4.30，and the degree of learning participation was 4.51. The feedback results of students showed that the clinical situation of the virtual simulation experiment was relatively close to the reality，with high comprehensive and repeatable，which could attract students’ attention，promote knowledge integration，and improve learning self-confidence. At the same time，students hoped to add the types of cases and targeted guidance to meet the personalized learning needs. Conclusion The virtual simulation experiment teaching design based on motivation theory can effectively stimulate students’ learning motivation.