Background Our 2012 review on therapeutic interventions for acute hamstring injuries found a lack of high-quality studies. The publication of new studies warranted an update. Objectives To update and reanalyse the efficacy of conservative treatments for hamstring injury. Data sources PubMed, EMBASE, Web of Science, Cochrane library, CINAHL and SPORTDiscus were searched till mid-February 2015. Study eligibility criteria Randomised controlled trials (RCTs) on the effect of conservative interventions versus a control group or other intervention for hamstring injuries (HI) were included. Data analysis The search results were screened independently by two authors. Risk of bias assessment was performed using a modified Downs and Black scale with a maximum score of 28. Meta-analysis was performed, where possible. Main results 10 RCTs (526 participants), including 6 new RCTs, were identified. Two RCTs were of good/ excellent quality, the rest were fair or poor (median Downs and Black score 16 (IQR 9)). Meta-analysis of two studies on rehabilitation (lengthening) exercises showed a significantly reduced time to return to play (HR 3.22 (95% CI 2.17 to 4.77), p<0.0001) but no difference in risk of re-injury. Meta-analysis of three studies investigating platelet-rich plasma (PRP) showed no effect when compared to control (HR 1.03 (95% CI 0.87 to 1.22), p=0.73). Limited evidence was found that progressive agility and trunk stability training may reduce re-injury rates. Conclusions Meta-analysis showed superior efficacy for rehabilitation exercises. PRP injection had no effect on acute hamstring injury. Limited evidence was found that agility and trunk stabilisation may reduce re-injury rates. The limitations identified in the majority of RCTs should improve the design of new hamstring RCTs.