Gilbert M Thomas P, Cuccui Jon, White William, Lynnerup Niels, Titball Richard W, Cooper Alan, Prentice Michael B
Henry Wellcome Ancient Biomolecules Centre, Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK.
Centre for Infection, Institute of Cell and Molecular Science, Barts and The London, Queen Mary's School of Medicine and Dentistry, 64 Turner St, London E1 2AD, UK.
Microbiology (Reading). 2004 Feb;150(Pt 2):341-354. doi: 10.1099/mic.0.26594-0.
This study reports the results of a collaborative study undertaken by two independent research groups to (a) confirm recent PCR-based detection of Yersinia pestis DNA in human teeth from medieval plague victims in France, and (b) to extend these observations over five different European burial sites believed to contain plague victims dating from the late 13th to 17th centuries. Several different sets of primers were used, including those previously documented to yield positive results on ancient DNA extracts. No Y. pestis DNA could be amplified from DNA extracted from 108 teeth belonging to 61 individuals, despite the amplification of numerous other bacterial DNA sequences. Several methods of extracting dentine prior to the DNA extraction were also compared. PCR for bacterial 16S rDNA indicated the presence of multiple bacterial species in 23 out of 27 teeth DNA extracts where dentine was extracted using previously described methods. In comparison, positive results were obtained from only five out of 44 teeth DNA extracts for which a novel contamination-minimizing embedding technique was used. Therefore, high levels of environmental bacterial DNA are present in DNA extracts where previously described methods of tooth manipulation are used. To conclude, the absence of Y. pestis-specific DNA in an exhaustive search using specimens from multiple putative European plague burial sites does not allow us to confirm the identification of Y. pestis as the aetiological agent of the Black Death and subsequent plagues. In addition, the utility of the published tooth-based ancient DNA technique used to diagnose fatal bacteraemias in historical epidemics still awaits independent corroboration.
本研究报告了两个独立研究小组开展的一项合作研究结果，目的是：（a）证实近期基于聚合酶链反应（PCR）对法国中世纪鼠疫受害者牙齿中鼠疫耶尔森菌DNA的检测；（b）将这些观察结果扩展至五个不同的欧洲埋葬地点，这些地点被认为包含可追溯至13世纪末至17世纪的鼠疫受害者。使用了几套不同的引物，包括先前记录在案的对古代DNA提取物产生阳性结果的引物。尽管扩增出了许多其他细菌DNA序列，但从61个人的108颗牙齿提取的DNA中未能扩增出鼠疫耶尔森菌DNA。还比较了在DNA提取之前提取牙本质的几种方法。针对细菌16S核糖体DNA（rDNA）的PCR表明，在27颗牙齿DNA提取物中的23份中存在多种细菌物种，这些提取物中的牙本质是使用先前描述的方法提取的。相比之下，在使用一种新型污染最小化包埋技术的44颗牙齿DNA提取物中，只有5份获得了阳性结果。因此，在使用先前描述的牙齿处理方法的DNA提取物中存在高水平的环境细菌DNA。总之，在对来自多个假定的欧洲鼠疫埋葬地点的标本进行详尽搜索时，未发现鼠疫耶尔森菌特异性DNA，这使我们无法确认鼠疫耶尔森菌是黑死病及后续鼠疫的病原体。此外，用于诊断历史流行病中致命菌血症的已发表的基于牙齿的古代DNA技术的实用性仍有待独立证实。
