OBJECTIVE To determine the effectiveness of the double-gloving method on preventing surgical glove perforation and blood contamination compared with single gloving. METHODS Systematic review. METHODS Seven electronic databases were searched including: Embase, CINAHL, OVID, Medline, Pubmed, Web of Science, and Foreign Medical Literature Retrieval Service in March 2020. METHODS Our systematic review and meta-analysis was reported following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) reporting guideline. Risk of bias of Cochrane Handbook (Version 5.1.0) was applied to evaluate the study quality. Revman 5.3 was used to calculate the effect size of odds ratio (OR) with 95% confidence interval (CI). Meta-analysis with forest plot and funnel plot was performed to compare the rate of surgical glove perforation and to determine the published bias, respectively. This review has been registered with ID: CRD42020189694 on the web site of PROSPERO. RESULTS Seven randomized controlled trials regarding the efficacy of double gloving on reducing surgical glove perforation were identified and a total of 7090 gloves were tested. After analyzing the pooled data, we identified that the rate of surgical glove perforation in the double-gloving group was lower than that of single gloving with statistical significance (OR = 0.75, 95% CI: 0.64-0.89, p < .05). It was statistically significant that surgical glove perforation was lower in the double-inner gloves as well as matched outer-inner perforated gloves compared with that of single glove (OR = 0.05, 95% CI: 0.03-0.07, p < .05). CONCLUSIONS Findings of this systematic review demonstrate that double gloving could reduce the rate of surgical-glove perforation. Meanwhile, the risk of being contaminated by a blood-borne pathogen during surgery could be reduced by wearing double gloves. We strongly suggest that surgical team members when operating should wear double gloves to protect themselves and reduce the risk of occupational blood exposure. CONCLUSIONS The necessity of double gloving for preventing blood contamination was demonstrated. The rate of surgical glove perforation is statistically significant in double-gloving group compared to single gloving. Double gloving could reduce the risk of being contaminated during surgery by blood-borne pathogen. Evidence is provided for surgical team and decision makers that double gloving could reduce occupational exposure.