1
Military Health Institute, Military Medical Agency, Tychonova 1, 160 01 Prague 6, Czech Republic
2
Institute of Molecular Genetics of the ASCR, v. v. i., Vídeňská 1083, 142 20 Prague 4, Czech Republic
3
National Museum, Department of Anthropology, Václavské náměstí 68, 115 79 Praha 1, Czech Republic
4
Faculty of Military Health Sciences, University of Defence, Třebešská 1575, 500 01 Hradec Králové, Czech Republic
5
Laboratory of Applied Proteome Analyses, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburská 89, 277 21 Liběchov, Czech Republic
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Genomics Core Facility, EMBL Heidelberg, Meyerhofstraße 1, 69117 Heidelberg, Germany
7
Institute of Pathology of the First Faculty of Medicine and General Teaching Hospital, Studničkova 2, 128 00 Prague, Czech Republic
8
Institute of Forensic Medicine and Toxicology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Studničkova 4, 128 21, Praha 2, Czech Republic
9
Institute of Organic Chemistry and Biochemistry of the CAS, Flemingovo náměstí 542/2, 166 10 Praha 6, Czech Republic
10
Military Institute of Forensic Medicine, Military University Hospital Prague, U Vojenské nemocnice 1200, 169 02 Praha 6
11
Bundeswehr Institute of Microbiology, Neuherbergstr. 11, 80937 Munich, Germany
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Abstract
Although smallpox has been known for centuries, the oldest available variola virus strains were isolated in the early 1940s. At that time, large regions of the world were already smallpox-free. Therefore, genetic information of these strains can represent only the very last fraction
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Although smallpox has been known for centuries, the oldest available variola virus strains were isolated in the early 1940s. At that time, large regions of the world were already smallpox-free. Therefore, genetic information of these strains can represent only the very last fraction of a long evolutionary process. Based on the genomes of 48 strains, two clades are differentiated: Clade 1 includes variants of variola major, and clade 2 includes West African and variola minor (Alastrim) strains. Recently, the genome of an almost 400-year-old Lithuanian mummy was determined, which fell basal to all currently sequenced strains of variola virus on phylogenetic trees. Here, we determined two complete variola virus genomes from human tissues kept in a museum in Prague dating back 60 and 160 years, respectively. Moreover, mass spectrometry-based proteomic, chemical, and microscopic examinations were performed. The 60-year-old specimen was most likely an importation from India, a country with endemic smallpox at that time. The genome of the 160-year-old specimen is related to clade 2 West African and variola minor strains. This sequence likely represents a new endemic European variant of variola virus circulating in the midst of the 19th century in Europe.
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