Detection of 400-year-old Yersinia pestis DNA in human dental pulp:: An approach to the diagnosis of ancient septicemia
Detection of 400-year-old Yersinia pestis DNA in human dental pulp:: An approach to the diagnosis of ancient septicemia
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DOI：
10.1073/pnas.95.21.12637
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发表时间：
1998-10-13
影响因子：
11.1
通讯作者：
Raoult, D
中科院分区：
文献类型：
--
作者：
Drancourt, M;Aboudharam, G;Raoult, D
据报道，古代败血症鼠疫流行病在两千年内造成数百万人死亡，然而，仅根据历史临床观察对古代败血症进行可靠的诊断是不可能的。由于缺乏合适的感染材料，无法直接证明古代败血症;因此，大多数感染如鼠疫的历史仍然是假设的。牙髓的耐久性，加上其天然的无菌性，使其成为一种合适的材料，以此为基础进行此类研究。我们假设它将成为鼠疫耶尔森氏菌的持久避难所。DNA提取物是从12颗未萌出牙齿的牙髓中制备的，这些牙齿是从16和18世纪法国被认为死于鼠疫的人的坟墓中挖掘出的骨骼中提取的（“鼠疫牙齿”），以及从7颗古代阴性对照牙齿中提取的。将古老的DNA提取物和引物特异性的人P-珠蛋白基因的PCR证明在这些制剂中的抑制剂的情况下。Y.鼠疫rpoB（RNA聚合酶P亚基编码基因）和公认的毒力相关pla（纤溶酶原激活物编码基因）重复产生的产物具有与现代细菌分离物不可区分的核苷酸序列。在12个鼠疫骨骼牙中有6个获得了特异性PLA序列，而在7个阴性对照中没有获得（P < 0.034，Fisher精确检验），对于历史上确定的受害者，实现了基于核酸的古代鼠疫确认，并且我们已经确认了该疾病在世纪末在法国的存在，牙髓是一个有吸引力的目标，在寻求确定病原学败血症疾病检测古代尸体。分子技术可以应用于这种材料，以解决历史爆发。
Ancient septicemic plague epidemics were reported to have killed millions of people for 2 millenniums, However, confident diagnosis of ancient septicemia solely on the basis of historical clinical observations is not possible. The lack of suitable infected material has prevented direct demonstration of ancient septicemia; thus, the history of most infections such as plague remains hypothetical. The durability of dental pulp, together with its natural sterility, makes it a suitable material on which to base such research, We hypothesized that it would be a lasting refuge for Yersinia pestis, the plague agent. DNA extracts were made from the dental pulp of 12 unerupted teeth extracted from skeletons excavated from 16th and 18th century French graves of persons thought to have died of plague ("plague teeth") and from 7 ancient negative control teeth. PCRs incorporating ancient DNA extracts and primers specific for the human P-globin gene demonstrated the absence of inhibitors in these preparations. The incorporation of primers specific for Y. pestis rpoB (the RNA polymerase P-subunit-encoding gene) and the recognized virulence-associated pla (the plasminogen activator encoding gene) repeatedly yielded products that had a nucleotide sequence indistinguishable from that of modern day isolates of the bacterium. The specific pla sequence was obtained from 6 of 12 plague skeleton teeth but 0 of 7 negative controls (P < 0.034, Fisher exact test), A nucleic acid-based confirmation of ancient plague was achieved for historically identified victims, and we have confirmed the presence of the disease at the end of 16th century in France, Dental pulp is an attractive target in the quest to determine the etiology of septicemic illnesses detected in ancient corpses. Molecular techniques could be applied to this material to resolve historical outbreaks.