Absence of Yersinia pestis-specific DNA in human teeth from five European excavations of putative plague victims
Absence of Yersinia pestis-specific DNA in human teeth from five European excavations of putative plague victims
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DOI：
10.1099/mic.0.26594-0
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发表时间：
2004-02-01
期刊：
影响因子：
2.8
通讯作者：
Prentice, MB
中科院分区：
文献类型：
--
作者：
Gilbert, MTP;Cuccui, J;Prentice, MB
这项研究报告了两个独立研究小组进行的一项合作研究的结果，以（a）确认最近在法国中世纪鼠疫受害者的人类牙齿中检测到鼠疫耶尔森氏菌DNA，（B）将这些观察结果扩展到五个不同的欧洲埋葬地点，这些地点被认为包含13世纪末至17世纪的鼠疫受害者。使用了几组不同的引物，包括以前记录的对古代DNA提取物产生阳性结果的引物。没有Y。鼠疫DNA可以从来自61个个体的108颗牙齿的DNA中扩增出来，尽管扩增了许多其他细菌的DNA序列。比较了几种在DNA提取前先提取牙本质的方法。细菌16S rDNA的PCR表明，在27个牙齿DNA提取物中的23个中存在多种细菌物种，其中使用先前描述的方法提取牙本质。相比之下，使用新型污染最小化包埋技术的44颗牙齿DNA提取物中只有5颗获得了阳性结果。因此，在使用先前描述的牙齿操作方法的DNA提取物中存在高水平的环境细菌DNA。总之，在使用多个假定的欧洲鼠疫埋葬地点的标本进行的详尽搜索中，Y鼠疫特异性DNA的缺失不允许我们确认Y鼠疫的鉴定。鼠疫是黑死病和随后的瘟疫的病原体。此外，已发表的基于牙齿的古代DNA技术在历史流行病中用于诊断致命菌血症的实用性仍有待独立证实。
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.