Dental pulp metagenomics reveals ancient human Herpesviridae.

Following a recent report on ancient human Herpesviridae (HHV), we adapted a standard protocol used for the high-throughput sequencing of modern DNA to recover ancient DNA (aDNA) from dental pulp specimens collected at two historical burial sites. Investigation of 36 soldiers buried in Sevastopol, Crimea, during the 1853-1856 Crimean War yielded genomic sequences covering 8.5% of the herpes simplex virus (HSV)-1 genome, 15.7% of the HSV-2 genome, 1.9% of the Epstein-Barr virus (EBV) genome and 3.9% of the cytomegalovirus genome. Further investigation of 16 civilians buried in the 17th century in Amiens, France, yielded genomic sequences covering 9.8% of the HHV-1 genome and 2.2% of the HHV-6A genome; all data obtained with parallel negative controls was appropriately negative. Specific qPCRs confirmed the presence of four HSV-1, two HSV-2 and one EBV sequences in Sevastopol and two HSV-1 and one HHV-6 sequences in Amiens. HHV-6 was further detected by fluorescent in situ hybridization. These results demonstrated the usefulness of modern metagenomic protocols for high-throughput sequencing of aDNA in dental pulp.