Healthcare-associated infections (HCAIs) cause significant morbidity and mortality in neonatal intensive care units (NICUs). Meticulous hand hygiene is the most effective strategy to prevent HCAI. However, hand hygiene compliance (HHC) is low, especially in ICUs. Hence, we aimed to evaluate the efficacy of strategies for improving HHC in NICUs. A systematic review of the literature and meta-analysis were carried out. PubMed, EMBASE, Cochrane CENTRAL and CINAHL were searched in October 2013. PRISMA guidelines were followed. The quality of included studies was assessed by the Newcastle-Ottawa scale (NOS). Sixteen eligible non-randomised studies were included. A total of 27,155 hand hygiene moments were observed. Meta-analysis using a random effects model indicated that a range of strategies, such as educational campaigns, musical parodies, reminders, easy access to hand hygiene sanitisers, UV sensors and performance feedback, improved HHC [odds ratio (OR) 2.04; 95 % confidence interval (CI) 1.40, 2.97]. Significant statistical heterogeneity was noted. Studies which specifically provided performance feedback at either the individual or group levels reported a more significant improvement in HHC compared to those that did not (OR 2.81; 95 % CI 1.32, 5.96 vs. OR 1.55; 95 % CI 1.13-2.11). Strategies to improve HHC in NICUs seem to be more effective when they include performance feedback at the personal or group levels. Randomised controlled trials (RCTs) specifically assessing the benefits of performance feedback in improving HHC are needed.