Dengue fever is currently a major public health problem in Burkina Faso. The aim of this study was to determine seroprevalence and dengue infection evolution during the years 2016 and 2017. The study population consisted of 3572 patients aged 0 to 84 years seen during general medicine consultation from January 2016 to November 2017 and referred to the laboratory of Saint Camille Hospital of Ouagadougou for serological diagnosis of dengue fever. The Dengue Duo Bioline SD Kit (Standard Diagnostic Inc., Korea) was used to detect the presence of dengue virus NS1 antigen and IgM/IgG antibodies in serum or plasma. An overall prevalence of 17.3% (617/3572) of NS1Ag was observed in our study population. Age groups 15 to 30 years were significantly more infected compared to children under 5 years of age. The peak of infection was between mid-October and mid-November corresponding to the end of the rainy season in Burkina Faso. The present study reports a high seroprevalence of acute dengue virus infection in symptomatic patients. The use of effective vector control strategies through the destruction of breeding sites, personal protection and enhanced surveillance from August is needed for dengue fever prevention in Burkina Faso.