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Background Surgical site infection (SSI) is one of the most frequent health care–associated infections. One of the practices to reduce their incidence is preoperative skin antisepsis. Two of the most commonly active components used are chlorhexidine gluconate and povidone iodine. Of 3 reviews conducted between 2010 and 2012 comparing antiseptics, 2 were in favor of chlorhexidine; however, the latest was unable to draw conclusions. Purpose To verify whether recent evidence supports the hypothesis that chlorhexidine in preoperative antisepsis is more efficient than other antiseptics in reducing SSI rates. Procedures We conducted a systematic review from 2000-2014 in all languages. The primary end point was SSI incidence and secondary skin bacterial colonization. Results Nineteen studies were included. Meta-analysis were conducted for comparable studies for both outcomes. The results of the meta-analysis, including all of the studies in which chlorhexidine was compared with iodophor, were in favor of chlorhexidine for both SSI incidence (risk ratio [RR], 0.70; 95% confidence interval [CI], 0.52-0.92) and bacterial skin colonization (RR, 0.45; 95% CI, 0.36-0.55). Conclusions There is moderate-quality evidence supporting the use of chlorhexidine for preoperative skin antisepsis and high-quality evidence that the use of chlorhexidine is associated with fewer positive skin cultures. Further rigorous trials will be welcomed to attain stronger evidence as to the best antiseptic to be used before surgery.