Purpose: Our aim was to evaluate the Microsoft Kinect sensor (MKS) as a markerless system for motion capture and analysis of lower limb motion, compare it with a state-of-the-art marker-based system (MBS), and investigate its accuracy in simultaneously capturing several lower limb joint movements on several planes while participants walked freely. Method: Participants were asked to walk while gait data were simultaneously recorded by both the MKS and the MBS. Software for analysing the Kinect data stream was developed using Microsoft Visual Studio and Kinect for Windows software development kits. Visual three-dimensional (3D) C-Motion software was used to calculate 3D joint angles of the MBS. Deviation of the joint angles calculated by the two systems was calculated using root-mean-square error (RMSE) on the basis of a designated formula. Results: The calculated RMSE average was <5° between the two systems, a level of difference that has practically no clinical significance. Conclusions: Quantitative measurements of the joint angles of the knee and hip can be acquired using one MKS with some accuracy. The system can be advantageous for clinical use, at the pre- and post-treatment stages of rehabilitation, at significantly lower costs. Further evaluation of the MKS should be performed with larger study populations.