In Senegal, during 2002–2007, 11 outbreaks of African swine fever (ASF) were reported to the World Organisation for Animal Health. Despite this, little was known of the epidemiology of ASF in the country. To determine the prevalence of ASF in Senegal in 2006, we tested serum specimens collected from a sample of pigs in the 3 main pig-farming regions for antibodies to ASF virus using an ELISA. Of 747 serum samples examined, 126 were positive for ASF, suggesting a prevalence of 16.9%. The estimated prevalences within each of the regions (Fatick, Kolda, and Ziguinchor) were 13.3%, 7.8%, and 22.1%, respectively, with statistical evidence to suggest that the prevalence in Ziguinchor was higher than in Fatick or Kolda. This regional difference is considered in relation to different farming systems and illegal trade with neighboring countries where the infection is endemic.