This study was performed to ascertain the effect on range of motion and muscle pain of dry needling of the gluteal muscles in athletes with hamstring pain of non- hamstring origin. 59 volunteer subjects with hamstring pain were recruited from a variety of sports (mostly Australian Rules Football). MRI scans of the hamstrings showed no hamstring pathology and hamstring pain was reproducible with pressure on the gluteal trigger points. Each subject underwent baseline measures of straight leg raise (SLR) and hip internal rotation (hip IR). They also recorded muscle pain and tightness at rest and on running activity on a Visual Analogue Scale (VAS). Subjects were randomised to a placebo or therapeutic needling group and received the intervention. Outcome measures were repeated immediately and at 24 and 72 hours. Results revealed no change in SLR or hip IR when compared to baseline measures. VAS scores for resting muscle sensations were essentially unchanged. Running hamstring pain and tightness and running gluteal pain and tightness improved significantly at the immediate post intervention measures (p = 0.001) and did not change significantly thereafter. There was no group effect on this result (p>0.05). Both placebo and therapeutic needling groups improved equally. In conclusion, both placebo and therapeutic dry needling had beneficial effects on subjective measures of hamstring discomfort, but not on straight leg raise or hip range of motion.