OpenPose-based motion analysis (OpenPose-MA), utilizing deep learning methods, has emerged as a compelling technique for estimating human motion. It addresses the drawbacks associated with conventional three-dimensional motion analysis (3D-MA) and human visual detection-based motion analysis (Human-MA), including costly equipment, time-consuming analysis, and restricted experimental settings. This study aims to assess the precision of OpenPose-MA in comparison to Human-MA, using 3D-MA as the reference standard. The study involved a cohort of 21 young and healthy adults. OpenPose-MA employed the OpenPose algorithm, a deep learning-based open-source two-dimensional (2D) pose estimation method. Human-MA was conducted by a skilled physiotherapist. The knee valgus angle during a drop vertical jump task was computed by OpenPose-MA and Human-MA using the same frontal-plane video image, with 3D-MA serving as the reference standard. Various metrics were utilized to assess the reproducibility, accuracy and similarity of the knee valgus angle between the different methods, including the intraclass correlation coefficient (ICC) (1, 3), mean absolute error (MAE), coefficient of multiple correlation (CMC) for waveform pattern similarity, and Pearson’s correlation coefficients (OpenPose-MA vs. 3D-MA, Human-MA vs. 3D-MA). Unpaired t-tests were conducted to compare MAEs and CMCs between OpenPose-MA and Human-MA. The ICCs (1,3) for OpenPose-MA, Human-MA, and 3D-MA demonstrated excellent reproducibility in the DVJ trial. No significant difference between OpenPose-MA and Human-MA was observed in terms of the MAEs (OpenPose: 2.4° [95%CI: 1.9-3.0°], Human: 3.2° [95%CI: 2.1-4.4°]) or CMCs (OpenPose: 0.83 [range: 0.99-0.53], Human: 0.87 [range: 0.24-0.98]) of knee valgus angles. The Pearson’s correlation coefficients of OpenPose-MA and Human-MA relative to that of 3D-MA were 0.97 and 0.98, respectively. This study demonstrated that OpenPose-MA achieved satisfactory reproducibility, accuracy and exhibited waveform similarity comparable to 3D-MA, similar to Human-MA. Both OpenPose-MA and Human-MA showed a strong correlation with 3D-MA in terms of knee valgus angle excursion.