In order to realize the precise positioning of radiation patients and the accurate measurement of tumor location in the process of precise radiation therapy, a medical robot positioning system based on binocular vision is established. First, the medical robot positioning measurement system is developed based on binocular vision; Secondly, based on the Tsai camera model and the influence of lens distortion, a new camera calibration method based on planar and three-dimensional checkerboard calibration templates is proposed to get the camera calibration parameters; Thirdly, the center point positions of markers are identified and their three-dimensional coordinates are calculated based on the robert gradient operator method for image segmentation. Finally, the measurement errors of all markers by binocular vision and the three-dimensional coordinate measuring instrument are used to achieve the position-validating for radiotherapy patients. The experimental results show that the camera calibration accuracy is 0.0365 mm and three-dimensional coordinates' average deviations of each marker are δX=0.573 mm, δY=0.495 mm, δZ=0.430 mm. The measurement method can improve the accuracy of calibration and positioning, and meet the clinical requirements for high-precision positioning system in precise radiotherapy.