Objective To quantify operation characteristics of the ankle rotating-traction-poking manipulation (RTPM) for treating acute lateral ankle sprain by using motion capture technology, so as to provide objective references for standardized operation of RTPM and its education inheritance. Methods A professional physiotherapist performed the RTPM on 60 volunteers with acute lateral ankle sprain. Motion capture system was used to acquire effective kinematic data during the RTPM, so as to make analysis and summarize rules. Results The average time of ankle rotating for six circles was 11.36 s and the average time of ankle traction and poking was 3.42 s. The average displacement of ankle traction was 36.94 mm and the average displacement of ankle poking was 22.44 mm. The average angle of ankle traction was 23.27°, and the average angle of ankle poking was 22.76°. During the RTPM for treating acute lateral ankle sprain, the average linear velocity of ankle rotating was 58.28 mm/s, and the average linear velocity of ankle traction and poking was 23.81 mm/s. The linear acceleration of ankle rotating was 0.43 mm/s2, and the linear acceleration of ankle traction and poking was 0.54 mm/s2. Conclusions The RTPM can be applied in clinical practice. During the RTMP, the principle of gentleness, rhythmicity and continuity should be followed. Under the premise of following physiological characteristics of ankle joint, treatment of the sprained ankle should be carried out with slow and uniform speed continuously.