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