Abstract Background: Venipuncture is most distressing and painful needle-related procedure experienced by pediatric where distraction combined with age-appropriate could be reduce procedural pain and anxiety among children. Virtual Reality (VR) is new technique for distraction and powerful technology implemented in a healthcare setting. Objective: Guided by Gate Control Theory, the study aimed to evaluate the effect of VR intervention on the pain and anxiety levels of children during venipuncture procedure. Method: Children were assigned randomly to one of two groups: a control group receiving standard care and VR group receiving VR distraction plus standard care during procedure. Pain scores were assessed using the Wong-Baker Faces Pain Scale (W-BFPS) and the Visual Analogue Scale (VAS) and the state of anxiety was measured by Facial Affective Scale (FAS). The anxiety and pain levels were evaluated based on the self-report approach from participated children. Results: A total of 144 children were randomly assigned to VR and control groups (72 VR and 72 control). The baseline characteristics of the VR and control groups were comparable. The self-reported W-BFPS and VAS revealed that pain level felt by the VR group during venipuncture procedure was lower than the level of pain indicated by control group significant at P<0.031; P<0.040 respectively. The self-reported Facial Affective Scale for anxiety level showed a significant difference between the VR and control groups (P < .001). Conclusion: It was determined that using the (VR)headset significantly reduced the pediatric pain felt and level of anxiety during a venipuncture procedure.