Objective: To compare surgical site infection rates in clean surgery before and after implementation of the checklist proposed by the World Health Organization. Methods: Observational, descriptive, retrospective correlational study performed in a general hospital. Sample consisting of 15,319 records of clean surgeries monitored by the hospital Infection Prevention and Control Service, in the trauma, orthopedics, cardiovascular, plastic, general, and urology specialties. Before implementing the checklist, 5,481 records were evaluated; 9,838 records were reviewed after. Analysis was performed with SPSS 22.0 software and application of the Pearson's chi-square test, considering p <0.05. Results: The overall infection rate in clean surgery was 4.17% in the pre-implementation period of the checklist and 1.10% post-implementation (p<0.05), with statistically significant reductions in spine, aneurysm and bypass, abdominoplasty, mammoplasty, herniorrhaphy and prostatectomy surgeries. Conclusion: A significant reduction was identified in the rate of surgical site infection in clean surgeries when comparing the pre- and post-implementation periods of the checklist proposed by the World Health Organization.