Molecular analysis of Mycobacterium tuberculosis DNA from a family of 18th century Hungarians
Molecular analysis of Mycobacterium tuberculosis DNA from a family of 18th century Hungarians
复制标题
DOI：
10.1099/mic.0.25961-0
复制
发表时间：
2003-01-01
期刊：
影响因子：
2.8
通讯作者：
Spigelman, M
中科院分区：
文献类型：
--
作者：
Fletcher, HA;Donoghue, HD;Spigelman, M
在18世纪匈牙利墓穴中发现的一位母亲和两个女儿的自然木乃伊遗骸被分析，使用多分子基因技术来研究结核病的流行病学和进化。从结核分枝杆菌基因组上的多个靶点中扩增出DNA，包括IS6110、gyrA、katG密码子463、oxR、Dna A-dna N、mtp40、plcD和直接重复序列(DR)。通过katG和gyrA基因分型、oxR和mtp40基因座的聚合酶链式反应(PCR)和基因分型，木乃伊遗骸中的菌株被鉴定为结核分枝杆菌，而不是牛分枝杆菌。Spoligo分型将样本分为两种菌株类型，在MT1801-plcD区域进行缺失筛查后，最初将菌株分为三种类型。然而，进一步的研究表明，明显的缺失是由于DNA保存不佳所致。通过比较扩增靶大小对扩增产物产量的影响，发现18世纪的结核分枝杆菌DNA与现代的结核分枝杆菌DNA有明显的差异。使用双中心系统来确认这项研究的结果，这清楚地表明了利用分子基因技术研究结核病历史病例的价值以及得出结论所需的注意事项。基因分型和分型结果与现代结核病流行的进化和传播的最新理论一致。
The naturally mummified remains of a mother and two daughters found in an 18th century Hungarian crypt were analysed, using multiple molecular genetic techniques to examine the epidemiology and evolution of tuberculosis. DNA was amplified from a number of targets on the Mycobacterium tuberculosis genome, including DNA from IS6110, gyrA, katG codon 463, oxyR, dnaA-dnaN, mtp40, plcD and the direct repeat (DR) region. The strains present in the mummified remains were identified as M. tuberculosis and not Mycobacterium bovis, from katG and gyrA genotyping, PCR from the oxyR and mtp40 loci, and spoligotyping. Spoligotyping divided the samples into two strain types, and screening for a deletion in the MT1801-plcD region initially divided the strains into three types. Further investigation showed, however, that an apparent deletion was due to poor DNA preservation. By comparing the effect of PCR target size on the yield of amplicon, a clear difference was shown between 18th century and modern M. tuberculosis DNA. A two-centre system was used to confirm the findings of this study, which clearly demonstrate the value of using molecular genetic techniques to study historical cases of tuberculosis and the care required in drawing conclusions. The genotyping and spoligotyping results are consistent with the most recent theory of the evolution and spread of the modern tuberculosis epidemic.