An accurate understanding of geographic and temporal seroprevalence trends is vital to reduce the public health impact of common flavivirus infections, including zika virus, dengue virus and Japanese encephalitis virus. Laboratory assays currently in use to examine the seroprevalence of these pathogens suffer from cross-reactivity and may mis-represent true seroprevalence. For technical and other reasons, our knowledge of flavivirus seroepidemiology in Thailand remains incomplete. This study offers an opportunity to examine historical and current flavivirus serodistribution, and will contribute to laboratory capacity within the Department of Disease Control, Ministry of Public Health.