Gait symmetry is fundamental to efficient locomotion and injury prevention. Recent advances in virtual reality (VR) technology offer immersive environments for gait analysis. However, the specific effects of VR on gait symmetry remain underexplored. This study aimed to investigate gait symmetry in healthy adults walking in a VR setting compared to a real-world laboratory, using statistical parametric mapping (SPM) to analyze each participant’s gait cycle. A dataset of 20 healthy adults was analyzed. Each participant performed multiple walking trials in both a real-world laboratory environment (RLab) and a corresponding virtual environment (VRLab). Kinematic data for the hip, knee, and ankle joints were recorded. SPM was then conducted on each participant’s data to detect asymmetries in the sagittal plane by comparing right and left joint angles within and between conditions. Significant asymmetry patterns were observed at the hip, knee, and ankle joints in both RLab and VRLab, but the magnitude and timing of these asymmetries varied across individuals. The hip joint exhibited relatively stable asymmetries between conditions, while the knee and ankle joints showed greater variability in VR, suggesting increased sensitivity to altered sensory inputs. VR environments can induce measurable changes in gait symmetry among healthy adults. These findings highlight VR’s potential as both a research tool and a rehabilitative platform, while also underscoring the need to address individual neuromuscular strategies and the sensory challenges inherent in immersive technology. Future studies should examine clinical populations and explore multi-planar kinematics to expand on these preliminary results.