Molecular Identification by "Suicide PCR" of Yersinia pestis as the Agent of Medieval Black Death
Medieval Black Death is believed to have killed up to one-third of the Western European population during the 14th century. It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pesti...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2000-11, Vol.97 (23), p.12800-12803 |
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creator | Raoult, Didier Aboudharam, Gerard Crubezy, Eric Larrouy, Georges Ludes, Bertrand Drancourt, Michel |
description | Medieval Black Death is believed to have killed up to one-third of the Western European population during the 14th century. It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer--its specificity is absolute. We believe that we can end the controversy: Medieval Black Death was plague. |
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It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer--its specificity is absolute. We believe that we can end the controversy: Medieval Black Death was plague.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.220225197</identifier><identifier>PMID: 11058154</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Adult ; Base Sequence ; Biological Sciences ; Child ; Children ; Dental Pulp - microbiology ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - analysis ; Epidemics ; Female ; France - epidemiology ; Graves ; History, 15th Century ; History, 16th Century ; History, 17th Century ; History, Medieval ; Humans ; Male ; Microbiology ; Middle Ages ; Molecular Sequence Data ; pla gene ; Plague ; Plague - epidemiology ; Plague - history ; Plague - microbiology ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Skeleton ; Suicide ; Teeth ; Tooth pulp ; Yersinia pestis ; Yersinia pestis - genetics ; Yersinia pestis - isolation & purification</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2000-11, Vol.97 (23), p.12800-12803</ispartof><rights>Copyright 1993-2000 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Nov 7, 2000</rights><rights>Copyright © 2000, The National Academy of Sciences 2000</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c519t-3103a1a6a0af36f682f1f5c9a2a9826f842cc51110346b24cee7c03afec1852c3</citedby><cites>FETCH-LOGICAL-c519t-3103a1a6a0af36f682f1f5c9a2a9826f842cc51110346b24cee7c03afec1852c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/97/23.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/123930$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/123930$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,315,729,782,786,805,887,27931,27932,53798,53800,58024,58257</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11058154$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Raoult, Didier</creatorcontrib><creatorcontrib>Aboudharam, Gerard</creatorcontrib><creatorcontrib>Crubezy, Eric</creatorcontrib><creatorcontrib>Larrouy, Georges</creatorcontrib><creatorcontrib>Ludes, Bertrand</creatorcontrib><creatorcontrib>Drancourt, Michel</creatorcontrib><title>Molecular Identification by "Suicide PCR" of Yersinia pestis as the Agent of Medieval Black Death</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Medieval Black Death is believed to have killed up to one-third of the Western European population during the 14th century. It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer--its specificity is absolute. 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It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer--its specificity is absolute. We believe that we can end the controversy: Medieval Black Death was plague.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>11058154</pmid><doi>10.1073/pnas.220225197</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adult Base Sequence Biological Sciences Child Children Dental Pulp - microbiology Deoxyribonucleic acid DNA DNA, Bacterial - analysis Epidemics Female France - epidemiology Graves History, 15th Century History, 16th Century History, 17th Century History, Medieval Humans Male Microbiology Middle Ages Molecular Sequence Data pla gene Plague Plague - epidemiology Plague - history Plague - microbiology Polymerase chain reaction Polymerase Chain Reaction - methods Skeleton Suicide Teeth Tooth pulp Yersinia pestis Yersinia pestis - genetics Yersinia pestis - isolation & purification |
title | Molecular Identification by "Suicide PCR" of Yersinia pestis as the Agent of Medieval Black Death |
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