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      High Throughput, Multiplexed Pathogen Detection Authenticates Plague Waves in Medieval Venice, Italy

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          Abstract

          Background

          Historical records suggest that multiple burial sites from the 14th–16 th centuries in Venice, Italy, were used during the Black Death and subsequent plague epidemics.

          Methodology/Principal Findings

          High throughput, multiplexed real-time PCR detected DNA of seven highly transmissible pathogens in 173 dental pulp specimens collected from 46 graves. Bartonella quintana DNA was identified in five (2.9%) samples, including three from the 16th century and two from the 15th century, and Yersinia pestis DNA was detected in three (1.7%) samples, including two from the 14th century and one from the 16th century. Partial glpD gene sequencing indicated that the detected Y. pestis was the Orientalis biotype.

          Conclusions

          These data document for the first time successive plague epidemics in the medieval European city where quarantine was first instituted in the 14th century.

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          Most cited references 54

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          Yersinia pestis--etiologic agent of plague.

          Plague is a widespread zoonotic disease that is caused by Yersinia pestis and has had devastating effects on the human population throughout history. Disappearance of the disease is unlikely due to the wide range of mammalian hosts and their attendant fleas. The flea/rodent life cycle of Y. pestis, a gram-negative obligate pathogen, exposes it to very different environmental conditions and has resulted in some novel traits facilitating transmission and infection. Studies characterizing virulence determinants of Y. pestis have identified novel mechanisms for overcoming host defenses. Regulatory systems controlling the expression of some of these virulence factors have proven quite complex. These areas of research have provide new insights into the host-parasite relationship. This review will update our present understanding of the history, etiology, epidemiology, clinical aspects, and public health issues of plague.
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            Natural history of plague: perspectives from more than a century of research.

            For more than a century, scientists have investigated the natural history of plague, a highly fatal disease caused by infection with the gram-negative bacterium Yersinia pestis. Among their most important discoveries were the zoonotic nature of the disease and that plague exists in natural cycles involving transmission between rodent hosts and flea vectors. Other significant findings include those on the evolution of Y. pestis; geographic variation among plague strains; the dynamics and maintenance of transmission cycles; mechanisms by which fleas transmit Y. pestis; resistance and susceptibility among plague hosts; the structure and typology of natural foci; and how landscape features influence the focality, maintenance, and spread of the disease. The knowledge gained from these studies is essential for the development of effective prevention and control strategies.
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              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 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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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2011
                10 March 2011
                : 6
                : 3
                Affiliations
                [1 ]Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), UMR CNRS 6236 IRD 198, IFR48, Faculté de Médecine, Université de la Méditerranée, Marseille, France
                [2 ]Anthropologie Bioculturelle, UMR 6578 CNRS, EFS, Université de la Méditerranée, Marseille, France
                [3 ]Soprintendenza Archeologica del Veneto, Venice, Italy
                University of Iowa, United States of America
                Author notes

                Conceived and designed the experiments: DR MD GA. Performed the experiments: TT LF MS. Analyzed the data: MS LF GA DR MD. Contributed reagents/materials/analysis tools: LF DR. Wrote the paper: TT MS MD GA.

                Article
                PONE-D-10-04467
                10.1371/journal.pone.0016735
                3053355
                21423736
                Tran et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                Page count
                Pages: 5
                Categories
                Research Article
                Medicine
                Infectious Diseases
                Bacterial Diseases
                Bubonic Plague
                Plagues
                Septicemic Plagues

                Uncategorized

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