BACKGROUND: The role of the operating room’s ventilation system in the prevention of surgical site infections (SSIs) is widely discussed, and existing guidelines do not reflect current evidence. In this context, laminar airflow ventilation was compared with conventional ventilation to assess their effectiveness in reducing the risk of SSIs. METHODS: We searched MEDLINE, Embase, Cochrane Central Register of Controlled Trials, and WHO regional medical databases from Jan 1, 1990, to Jan 31, 2014. We updated the search for MEDLINE for the period between Feb 1, 2014, and May 25, 2016. We included studies most relevant to our predefined question: is the use of laminar airflow in the operating room associated with the reduction of overall or deep SSI as outcomes in patients of any age undergoing surgical operations? We excluded studies not relevant to the study question, studies not in the selected languages, studies published before Jan 1, 1990, or after May 25, 2016, meeting or conference abstracts, and studies of which the full text was not available. Data were extracted by two independent investigators, with disagreements resolved through further discussion. Authors were contacted if the full-text article was not available, or if important data or information on the paper’s content was absent. Studies were assessed for publication bias. Grading of recommendations assessment, development, and evaluation was used to assess the quality of the identified evidence. Meta-analyses were done with RevMan (version 5.3). FINDINGS: We identified 1947 records of which 12 observational studies were comparing laminar airflow ventilation with conventional turbulent ventilation in orthopaedic, abdominal, and vascular surgery. The meta-analysis of eight cohort studies showed no difference in risk for deep SSIs following total hip arthroplasty (330â146 procedures, odds ratio [OR] 1Â·29, 95% CI 0Â·98-1Â·71; p=0Â·07, I(2)=83%). For total knee arthroplasty, the meta-analysis of six cohort studies showed no difference in risk for deep SSIs (134â368 procedures, OR 1Â·08, 95% CI 0Â·77-1Â·52; p=0Â·65, I(2)=71%). For abdominal and open vascular surgery, the meta-analysis of three cohort studies found no difference in risk for overall SSIs (63â472 procedures, OR 0Â·75, 95% CI 0Â·43-1Â·33; p=0Â·33, I(2)=95%). INTERPRETATION: The available evidence shows no benefit for laminar airflow compared with conventional turbulent ventilation of the operating room in reducing the risk of SSIs in total hip and knee arthroplasties, and abdominal surgery. Decision makers, medical and administrative, should not regard laminar airflow as a preventive measure to reduce the risk of SSIs. Consequently, this equipment should not be installed in new operating rooms. FUNDING: None.