In current domestic research on laparoscopic training, researchers usually consider instrument movement path in the hand-eye coordination relationship. However, they ignore the information contained in visual cues by which could guide and control instrument movements. Studies in other areas have shown that trainers can improve their perceptual-motor skills by gaze training. This paper was designed to examine the effectiveness of eye gaze tracking technology in laparoscopic training and to analyze gaze strategy of the subjects in different training methods. The Tobii X1 Light Eye Tracker was used to track the gaze position of subjects when they were performing the two-handed transferring task in box trainer, and to obtain parameters related to gaze strategy including the efficiency of task completion, as well as visual search, visual processing and observation transfer analysis based on Markov chain model. The results showed that the completion time during the last training in gaze training group was decreased by 101.5 s comparing to the first training. Compared with video training group, gaze strategy of gaze training group has a significant change, such as fixation and saccade duration rate was increased by 38%, fixation duration on target area was increased, and saccade amplitude increased by 0.58°, and the probability of the fixation point transferring to equipment decreased by 15%. The results demonstrated that eye gaze tracking technology can be used in laparoscopic training, and can improve the subjects’ skills and shorten the learning curve by learning gaze strategies of experts.