This prospective cohort study evaluated the relationship of hamstring and lumbar spine flexibility to hamstring injury. Sixty-seven senior male Australian Rules footballers were videotaped while performing a toe-touch test from erect standing. The Peak Motion Measurement System was used to obtain measurements of end range hip flexion, lumbar flexion, toe-touch distance (TTD) and the ratio of lumbar spine flexion to hip flexion. Over the following football season, eight subjects (11.9 per cent) sustained a hamstring strain. Results showed no significant difference between the hamstring injured or uninjured players for any of the measured variables with no variable able to predict the likelihood of injury (p >0.05). In this cohort, the toe-touch test would not appear to be a useful screening tool to identify footballers at risk for hamstring strain.