Background: Virtual reality serves as a promising intervention for alleviating pain and anxiety in children. Aim: To evaluate the effectiveness of a virtual reality application on mitigating children's pain and anxiety throughout phlebotomy. This study employed a quasi-experimental research design. The investigation was conducted at the outpatient clinics and emergency pediatric department of Assiut University Children Hospital. A purposive sample of one hundred child was included in the study. The instruments were a structured interview questionnaire for children, a pain rating scale, and the Beck Anxiety Inventory scale. The results demonstrated statistically significant differences in pain and anxiety levels between the posttest and pretest. Moreover, children utilizing a virtual reality exhibited reduced pain and anxiety compared to those receiving conventional hospital treatment. Conclusion: The virtual reality application positively influenced pain and anxiety levels during phlebotomy. Consequently, it was Recommended that virtual reality applications be employed as conventional therapies for managing pain and anxiety during pediatric phlebotomy in clinical environments.