No sequences of genotype 1, the currently circulating virus, were detected. The five samples positive with only VP-qPCR showed 93.7–99.2% identity to genotype 2 (AJ717293). Among the samples positive with both qPCRs (n = 43), 41 showed 96.4%–99.6% total nucleotide identity to the B19V genotype 2 reference sequence (GenBank accession number AJ717293), while the remaining two samples (#12 and #38) showed 98.0% and 97.8% identity to genotype 3 (GenBank accession number AY083234). The product of the VP-qPCR is shorter (121 bp vs 154 bp) and may therefore amplify more effectively the possibly fragmented DNA sequences in these bone samples.Īll B19V qPCR products were sequenced. The viral loads of the five samples positive solely by the VP-qPCR ranged from 2.7 × 10 0 to 1.7 × 10 3/1 μg of total DNA, thus the difference was not due to copy number or a lack of Pan-B19V qPCR sensitivity.
For this purpose, two quantitative PCRs (qPCR), Pan-B19V qPCR and VP-qPCR, targeting distinct conserved areas of the viral genome (non-structural and viral protein respectively), were used.ī19V DNA was detected in 48 samples (45%), of which 43 were positive by both qPCRs and five by the VP-qPCR only. The B19V genomic prevalence was determined in DNA extracts of long bones of 106 anonymous World War II casualties. The present work not only explores the suitability of bone for search of viral DNA but also reveals unambiguously the forms of B19V that circulated in the first half of the 20 th century. In Northern Europe, both genotypes 1 and 2 have been encountered in soft tissues of elderly individuals 7, 8, 17 yet a clear perspective on the endemic prevalence of each type across the years is lacking, as the time of primary infection is not known. Of these, the most extensively studied and the one responsible for most current clinical cases is genotype 1. The virus, however, replicates in erythroid progenitor cells in the bone marrow 13, 14, 15 and also has been found in mesenchymal stromal cells, which can differentiate into cartilage and bone 16.ī19V has three genotypes that are differently distributed around the globe. B19V DNA has been detected in a range of tissues and organs 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 but as of yet there is no direct evidence of its persistence in bone.
The remains have over the past 17 years been searched for and repatriated upon discovery to Finland for DNA-based identification 1.Īs a proof of principle, we examined these bones for human parvovirus B19 (B19V), a highly prevalent DNA virus establishing lifelong tissue persistence.
In the present study we searched for viral DNA skeletal remains of putative Finnish soldiers that went missing in action during the Second World War (WWII) and whose bodies had been decaying in the boreal forest in current Russian territory ever since. Thus, tracing the footprints of viruses in human remains may reveal important clues on their distribution, adaptation and on the influence that they may have on us.ĭNA is most likely to be preserved across time in bone yet little is known about viral persistence in this organ. Understanding viral evolutionary dynamics may be crucial for better monitoring and predicting the epidemiological trajectory of clinically relevant viruses. The advent of new genomic and sequencing technologies has uncovered in recent years a myriad of viral sequences that were previously unknown to coexist with humans.
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