PCR diagnostics of Mycobacterium tuberculosis in historic human long bone remains from 18th century burials in Kaiserebersdorf, Austria

BMC Research Notes, Sep 2008

Background In the present pilot study we applied recently published protocols for detecting Mycobacterium tuberculosis in human remains. We screened long bones from an 18th century cemetery and skulls from the anatomical "Weisbach collection" (19th century). In addition, besides the study of abundance of tuberculosis in inmates of the poorhouse itself, we were interested to test whether in this particular instance tuberculosis can be identified from cortical bones, which are rarely affected by tuberculosis, but mostly better preserved than the vertebral bodies or epiphyses. Method The DNA extractions from the bone samples were obtained following established ancient DNA protocols. Subsequently extracts were subjected to a series of PCR amplifications using primer pairs published previously [1, 2]. PCR products of the expected size were subsequently sequenced. Results Only primers targeting the repetitive IS6110 insertion sequence yielded PCR products of appropriate size. In one sample only (skull sample WB354 of the "Weisbach collection") sequence analysis revealed an authentic M. tuberculosis sequence that matched to a reference sequence from GenBank. Conclusion With a variety of established PCR approaches we failed to detect M. tuberculosis DNA in historic human femurs from an 18th century cemetery relating to a poor house in Kaiserebersdorf, Austria. Our data may indicate that in this particular case, thoracic or lumbar vertebrae, i.e. bones that are severely affected by the disease, would be more suitable for molecular diagnostics than long bones. However, the unpredictable state of DNA preservation in bones from museum collections does not allow any general recommendation of any type of bone.

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PCR diagnostics of Mycobacterium tuberculosis in historic human long bone remains from 18th century burials in Kaiserebersdorf, Austria

BMC Research Notes Short Report PCR diagnostics of Mycobacterium tuberculosis in historic human long bone remains from 18th century burials in Kaiserebersdorf, Austria Lutz Bachmann 2 Barbara Däubl 1 Charlotte Lindqvist 2 3 Luise Kruckenhauser 1 Maria Teschler-Nicola 0 Elisabeth Haring 1 0 Museum of Natural History Vienna, Department of Anthropology , Burgring 7, A-1010, Vienna , Austria 1 Museum of Natural History Vienna , 1st Zoology. Department, Molecular Systematics, Burgring 7, A-1010, Vienna , Austria 2 Natural History Museum, Department for Zoology, University of Oslo , PO Box 1172, Blindern, NO-0318, Oslo , Norway 3 Department of Biological Sciences, The State University of New York at Buffalo , Buffalo, NY 14260 , USA Background: In the present pilot study we applied recently published protocols for detecting Mycobacterium tuberculosis in human remains. We screened long bones from an 18th century cemetery and skulls from the anatomical "Weisbach collection" (19th century). In addition, besides the study of abundance of tuberculosis in inmates of the poorhouse itself, we were interested to test whether in this particular instance tuberculosis can be identified from cortical bones, which are rarely affected by tuberculosis, but mostly better preserved than the vertebral bodies or epiphyses. Method: The DNA extractions from the bone samples were obtained following established ancient DNA protocols. Subsequently extracts were subjected to a series of PCR amplifications using primer pairs published previously [1,2]. PCR products of the expected size were subsequently sequenced. Results: Only primers targeting the repetitive IS6110 insertion sequence yielded PCR products of appropriate size. In one sample only (skull sample WB354 of the "Weisbach collection") sequence analysis revealed an authentic M. tuberculosis sequence that matched to a reference sequence from GenBank. Conclusion: With a variety of established PCR approaches we failed to detect M. tuberculosis DNA in historic human femurs from an 18th century cemetery relating to a poor house in Kaiserebersdorf, Austria. Our data may indicate that in this particular case, thoracic or lumbar vertebrae, i.e. bones that are severely affected by the disease, would be more suitable for molecular diagnostics than long bones. However, the unpredictable state of DNA preservation in bones from museum collections does not allow any general recommendation of any type of bone. - Background Bone tuberculosis (Spondylitis tuberculosa, "Pott's disease") and joint tuberculosis are caused by Mycobacterium tuberculosis and appear among others as late manifestations of a tuberculosis infection. Because of the inflammatory bony changes, e.g., partial or total destruction of the vertebral bodies or joint elements, manifestations of tuberculosis infections of (pre)historic human skeletal remains are very often identifiable by means of macroscopic inspections. Since the spread of M. tuberculosis depends on population density, its verification is also of high interest for the reconstruction of population dynamic processes in archaeology and anthropology [3]. Nevertheless, post mortem destructions with substantial substance loss or other pathologies with a similar appearance such as, e.g., destruction and remodeling of joint surfaces caused by a fracture and dislocation of a joint element, or an idiopathic avascular necrosis of the femoral head, may lead occasionally to erroneous diagnoses, which may miss- or under-represent the prevalence of tuberculosis in past human populations. An unambiguous molecular identification of tuberculosis for historic human bone remains has therefore been highly appreciated by anthropologists and paleo-epidemiologists. In recent years, tuberculosis has been diagnosed from a variety of historic human bone remains using ancient DNA methodology. Spigelman and Lemma [4] were the first to detect authentic DNA of M. tuberculosis in pre-European-contact human remains from Borneo by means of PCR amplification. This study has been criticized to some extent for technical issues, but the results were confirmed some ten years later [5]. Other examples of detecting M. tuberculosis in historic samples include ancient Egyptian mummies [6] and twelve approximately 140 – 1,200 years old mummies excavated from the Andes Mountain region of South America [1]. In seven samples Mycobacterium DNA could be detected out of which two proved positive for M. tuberculosis. In another study several loci of the M. tuberculosis genome were targeted in DNA extracted from naturally mummified remains from three 18th century individuals from Hungary by means of PCR and spoligotyping [7], and M. tuberculosis rather than Mycobacterium bovis was identified as the cause of the disease. Taylor et al. [2] were able to confirm the morphology based diagnosis of tuberculosis for approximately 2,200 years old human remains excavated from Tarrant Hinton, Dorset, United Kingdom, (...truncated)


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Lutz Bachmann, Barbara Däubl, Charlotte Lindqvist, Luise Kruckenhauser, Maria Teschler-Nicola, Elisabeth Haring. PCR diagnostics of Mycobacterium tuberculosis in historic human long bone remains from 18th century burials in Kaiserebersdorf, Austria, BMC Research Notes, 2008, pp. 83, Volume 1, Issue 1, DOI: 10.1186/1756-0500-1-83