BACKGROUND: The hamstrings remain the most injured muscle group within the Australian Football League (AFL). OBJECTIVE: To investigate preseason measures of hip and knee joint position sense (JPS) and prospective hamstring injury in AFL players. DESIGN: Prospective cohort study. METHODS: A total of 116 AFL players were recruited for this study. JPS was assessed with 3-D sensors using mono-articular hip (45° flexion and 0°) and knee (90° and 45° flexion) joint reproduction tests conducted in the preseason. Hamstring injury data were collected prospectively in the following AFL season. Wilcoxon-signed rank tests were used to assess between the subsequently injured and uninjured limbs. Mann-Whitney U tests were used to assess between group differences and odds ratio (OR) were used to predict players at risk of hamstring injury. RESULTS: Eight players with JPS data sustained a season hamstring injury and 108 players did not. JPS was not significantly different between the subsequently injured and uninjured limbs (all P values > 0.05). No significant differences for any JPS measure were found between the subsequently injured and uninjured players (all p's > 0.05). ORs did not achieve significance for AE (2.7, p = 0.21) or for RMSE (OR = 1.9, p = 0.44). CONCLUSION: Lower limb JPS measures were not predictive of hamstring injury in AFL players.