Skin-mounted marker based motion capture systems are widely used in measuring the movement of human joints. Kinematic measurements associated with skin-mounted markers are subject to soft tissue artifacts (STA), since the markers follow skin movement, thus generating errors when used to represent motions of underlying bone segments. We present a novel ultrasound tracking system that is capable of directly measuring tibial and femoral bone surfaces during dynamic motions, and subsequently measuring six-degree-of-freedom (6-DOF) tibiofemoral kinematics. The aim of this study is to quantitatively compare the accuracy of tibiofemoral kinematics estimated by the ultrasound tracking system and by a conventional skin-mounted marker based motion capture system in a cadaveric experimental scenario. Two typical tibiofemoral joint models (spherical and hinge models) were used to derive relevant kinematic outcomes. Intra-cortical bone pins equipped with optical markers were inserted in the tibial and femoral bones to serve as a reference to provide ground truth kinematics. The ultrasound tracking system resulted in lower kinematic errors than the skin-mounted markers (the ultrasound tracking system: maximum root-mean-square (RMS) error 3.44° for rotations and 4.88 mm for translations, skin-mounted markers with the spherical joint model: 6.32° and 6.26 mm, the hinge model: 6.38° and 6.52 mm). Our proposed ultrasound tracking system has the potential of measuring direct bone kinematics, thereby mitigating the influence and propagation of STA. Consequently, this technique could be considered as an alternative method for measuring 6-DOF tibiofemoral kinematics, which may be adopted in gait analysis and clinical practice.