Identification of viral DNA (Anelloviridae) in a 200-year-old dental pulp sample (Napoleon's Great Army, Kaliningrad, 1812).

Ancient human remains are potential sources of biological information including traces of past infections, since previous studies have demonstrated the effective detection of several bacterial agents or host-integrated viruses in old biological remnants like tissues or teeth. Studies of skeletal dental pulp samples are of particular interest since this location is potentially exposed to bloodborne agents during life through its vascularization, and could be considered as well preserved from environment after death of the host. DNA viruses belonging to the family Anelloviridae are highly present in human populations where they harbor an extreme genetic diversity but a yet undefined implication in hosts' health. We hypothesized that anelloviruses would be detected in ancient remains and that they may also serve as tracer viruses for the study of other viral agents. We analyzed 200-year-old dental pulp samples from remains of soldiers of Napoleon's Great Army during the Russian Retreat. Successful detection of Anelloviridae DNA by PCR was obtained for 1/21 ancient samples tested. The sequence identified showed 23% nucleotide divergence with the closest group of modern isolates (genus Gammatorquevirus), and was confirmed as phylogenetically distinct from those identified in saliva samples from the two investigators in charge of the study (genera Alphatorquevirus and Betatorquevirus). PCR directed toward the human beta globin gene was also performed. Negative controls were negative. Our results demonstrate that an ubiquitary, non-integrated, DNA virus is detectable from ancient biological material, with potential developments in terms of evolution studies or subsequent molecular investigations involving further viral agents.

[1]  R. Prŷs-Jones,et al.  Extracting DNA from museum bird eggs, and whole genome amplification of archive DNA , 2008, Molecular ecology resources.

[2]  S. Pääbo,et al.  Genetic analyses from ancient DNA. , 2004, Annual review of genetics.

[3]  H. Donoghue,et al.  Mycobacterium leprae DNA from ancient bone detected by PCR , 1994, The Lancet.

[4]  M. Spigelman,et al.  The use of the polymerase chain reaction (PCR) to detect Mycobacterium tuberculosis in ancient skeletons , 1993 .

[5]  H. Okamoto.  TT viruses in animals. , 2009, Current topics in microbiology and immunology.

[6]  P. de Micco,et al.  Circular genomes related to anelloviruses identified in human and animal samples by using a combined rolling-circle amplification/sequence-independent single primer amplification approach. , 2007, The Journal of general virology.

[7]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[8]  D. Raoult,et al.  Evidence for louse-transmitted diseases in soldiers of Napoleon's Grand Army in Vilnius. , 2006, The Journal of infectious diseases.

[9]  S. Horai,et al.  The presence of ancient human T-cell lymphotropic virus type I provirus DNA in an Andean mummy , 1999, Nature Medicine.

[10]  P. D. Micco,et al.  Les anellovirus (TTV et variants) : données actuelles dix ans après leur découverte , 2008 .

[11]  A. Geddes,et al.  The history of smallpox. , 2006, Clinics in dermatology.

[12]  M. Kudo,et al.  Hepatocellular carcinomas infected with the novel TT DNA virus lack viral integration. , 1998, Biochemical and biophysical research communications.

[13]  E. Rybicki,et al.  Experimental observations of rapid Maize streak virus evolution reveal a strand-specific nucleotide substitution bias , 2008, Virology Journal.

[14]  C. Millar,et al.  New developments in ancient genomics. , 2008, Trends in ecology & evolution.

[15]  E. Delwart.  Viral metagenomics , 2007, Reviews in medical virology.

[16]  J. Taubenberger,et al.  Origin and evolution of the 1918 "Spanish" influenza virus hemagglutinin gene. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  T. Nishizawa,et al.  A novel DNA virus (TTV) associated with elevated transaminase levels in posttransfusion hepatitis of unknown etiology. , 1997, Biochemical and biophysical research communications.

[18]  D. Raoult,et al.  Bartonella quintana in a 4000-year-old human tooth. , 2005, The Journal of infectious diseases.

[19]  S. Colowick,et al.  Methods in Enzymology , Vol , 1966 .

[20]  C. Mulligan.  Isolation and analysis of DNA from archaeological, clinical, and natural history specimens. , 2005, Methods in enzymology.

[21]  P. Biagini,et al.  Classification of TTV and related viruses (anelloviruses). , 2009, Current topics in microbiology and immunology.

[22]  P. Simmonds,et al.  Resistance of porcine circovirus and chicken anemia virus to virus inactivation procedures used for blood products , 2006, Transfusion.

[23]  D. Raoult,et al.  Molecular identification by "suicide PCR" of Yersinia pestis as the agent of medieval black death. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[25]  G. Fornaciari,et al.  Human papillomavirus in a 16th century mummy , 2003, The Lancet.

[26]  A. Nerlich,et al.  Molecular analysis of ancient microbial infections. , 2002, FEMS microbiology letters.

[27]  M. Prentice,et al.  Absence of Yersinia pestis-specific DNA in human teeth from five European excavations of putative plague victims. , 2004, Microbiology.

[28]  H. Okamoto.  History of discoveries and pathogenicity of TT viruses. , 2009, Current topics in microbiology and immunology.