BackgroundPaleomicrobiological data have clarified thatPlasmodiumspp. was circulating in the past in southern European populations, which are now devoid of malaria. The aim of this study was to evaluate the efficacy of immunodetection and, more particularly, rapid diagnostic tests (RDT), in order to further assessPlasmodiuminfections in ancient northern European populations.MethodsA commercially available RDT, PALUTOP®+ 4 OPTIMA, which is routinely used to detect malaria, was used to detectPlasmodiumantigens from proteins recovered from ancient specimens extracted from 39 dental pulp samples. These samples were collected from 39 individuals who were buried in the sixth century, near the site of the current Palace of Versailles in France. Positive and negative controls were also used. Antigens detected were quantified using chemiluminescence imaging system analysis.ResultsPlasmodiumantigens were detected in 14/39 (35.9%) individuals, includingPlasmodium vivaxantigens in 11 individuals andPlasmodium falciparumantigens co-detected in two individuals, while Pan-Plasmodiumantigens were detected in three individuals. Controls all yielded expected results.ConclusionsThe data reported here showed that RDTs are a suitable tool for detectingPlasmodiumspp. antigens in ancient dental pulp samples, and demonstrated the existence of malaria in Versailles, France, in the sixth century.Plasmodium vivax,which is regarded as being responsible for an attenuated form of malaria and less deadly forms, was the most prevalent species. This illustrates, for the first time in ancient populations, co-infection withP. falciparum, bringing into question the climate-driven ecosystems prevailing at that time in the Versailles area.