Detection of Yersinia pestis DNA in two early medieval skeletal finds from aschheim (Upper Bavaria, 6th century AD)
Detection of Yersinia pestis DNA in two early medieval skeletal finds from aschheim (Upper Bavaria, 6th century AD)
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DOI：
10.1002/ajpa.10276
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发表时间：
2005-01-01
影响因子：
2.8
通讯作者：
Grupe, G
中科院分区：
文献类型：
--
作者：
Wiechmann, I;Grupe, G
在对双重土葬进行分子遗传学调查的过程中，据推测是来自Aschheim（上巴伐利亚，公元世纪）的母子合葬，通过线粒体DNA、分子性别鉴定和核DNA多态性分析，检测到鼠疫耶尔森菌特异性DNA。对从一个骨骼的两颗牙齿和另一个骨骼的四颗牙齿中获得的DNA提取物进行了分子分析。引物对YP 12 D/YP 11 R（Raoult等人[2000] Proc.Natl. Acad. Sci. 97：12800-12803），能够扩增Y.鼠疫菌质粒pPCP 1 pla序列，产生预期片段大小的扩增产物。经BLACKS2.2.2分析，这些扩增产物的序列与现代Y. pestis pla基因序列，除一个扩增产物有单碱基替换外，其余均为阴性。使用独立引物对YP 11 D/YP 10 R的“自杀PCR”的应用（Raoult等人[2000] Proc.Natl. Acad. Sci. 97：12800-12803）导致扩增产物与现代Y. pestis pla基因序列。观察到的偏差可能是由于模板DNA中的错误编码病变。没有现代Y。鼠疫DNA被引入研究所，因此没有携带阳性对照沿着。所有浸提和PCR对照品均保持阴性。在这两具埋葬于公元世纪后半叶的骸骨中，构成了Y存在的分子支持证据。在有记录的第一次大流行期间，鼠疫是鼠疫的病原体。(C)2004威利-利斯公司
In the course of a molecular genetic investigation of a double inhumation, presumably a mother/ child burial from Aschheim (Upper Bavaria, 6th century A.D.), which included analysis of mitochondrial DNA, molecular sexing, and polymorphic nuclear DNA, Yersinia pestis-specific DNA was detected. Molecular analyses were performed on DNA extracts obtained from two teeth of one skeleton and four teeth of the other. The use of the primer pair YP12D/YP11R (Raoult et al. [2000] Proc. Natl. Acad. Sci. 97:12800-12803), able to amplify part of the Y. pestis plasmid pPCP1 pla sequence, resulted in amplification products of the expected fragment size. Using BLASTN 2.2.2, the sequences of these amplification products shared 100% identity with that of the modern Y. pestis pla sequence in GenBank, with the exception of one amplification product which revealed a single base substitution. The application of a "suicide PCR" with the independent primer pair YP11D/YP10R (Raoult et al. [2000] Proc. Natl. Acad. Sci. 97:12800-12803) resulted in amplification products which shared a 96-98% homology with that of the modern Y. pestis pla sequence in GenBank. The observed deviations were presumably due to miscoding lesions in the template DNA. No modern Y. pestis DNA was introduced into the institute, and thus no positive controls were carried along. All extraction and PCR controls remained negative. The identification of Y, pestis-specific DNA sequences in these two skeletons, buried in the second half of the 6th century A.D., constitutes molecularly supported evidence for the presence of Y. pestis, the causative agent of plague, during the first pandemic recorded. (C) 2004 Wiley-Liss, Inc.