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Summary: Objectives: Our study aimed to update the seroprevalence and factors associated with anti‐dengue virus (DENV) antibody positivity among blood donors and to discuss their implications for blood supply. Background: Questions on the potential transmission of DENV by transfusion increased after the documentation of the risk of transmission of the West Nile virus. This risk was estimated after transfusion of DENV RNA‐positive blood units of up to 37.5%. In Burkina Faso, very few studies on DENV in blood donors have been conducted. As a result, there were no reliable data on DENV to allow the implementation of appropriate measures to control the risk of transmission of the dengue virus by blood transfusion. Methods: We conducted a 4‐week cross‐sectional study from December 4 to 30, 2016. Blood donors of both genders, aged 18‐60 years, accepted for blood donation after medical selection were consecutively enrolled. Results: Our study included a total of 1007 blood donors, in which donors living in urban areas represented 78.2%. The mean age was 26.1 ± 8.1 years. After adjustment in a multiple regression logistic model, the odds of having IgG anti‐DENV increased as age increased. The odds of DENV was 53% lower in rural areas (OR = 0.47; P =.000) compared to urban settings and 42% lower in mobile sites (OR = 0.58; P =.03) compared to fixed ones. Conclusion: Our study provides new and useful insights for future research on the risk of TT‐DENV throughout blood transfusion. [ABSTRACT FROM AUTHOR]