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


METHODS
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 12. 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.


CONCLUSIONS
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.
Bachmann
Lutz
L
Natural History Museum, Department of Zoology, University of Oslo, PO Box 1172, Blindern, NO-0318, Oslo, Norway. bachmann@nhm.uio.no
Däubl
Barbara
B
Lindqvist
Charlotte
C
Kruckenhauser
Luise
L
Teschler-Nicola
Maria
M
Haring
Elisabeth
E
Am J Phys Anthropol. 2005 Jan;126(1):32-47
15386245
J Med Microbiol. 2001 Apr;50(4):355-366
11289521
Am J Phys Anthropol. 2002 Sep;119(1):27-36
12209571
Am J Clin Pathol. 1994 Jun;101(6):733-7
8209861
Nucleic Acids Res. 1993 Aug 11;21(16):3913-4
8396242
J Clin Microbiol. 2002 Dec;40(12):4738-40
12454182
Kansenshogaku Zasshi. 2003 Mar;77(3):146-9
12708007
Mol Cell Probes. 1991 Feb;5(1):73-80
1673228
J Clin Microbiol. 1993 Aug;31(8):1987-95
7690367
J Infect Dis. 1990 May;161(5):977-81
2109022
Lancet. 1997 Nov 8;350(9088):1404
9365482
Microbiology (Reading). 2003 Jan;149(Pt 1):143-51
12576588
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3684-9
11891304
Microbiology (Reading). 2007 Apr;153(Pt 4):1243-1249
17379733
Lancet Infect Dis. 2004 Sep;4(9):584-92
15336226
Mol Biol Evol. 2004 May;21(5):957-64
15014140
Forensic Sci Int. 2008 May 2;177(1):32-6
18063334
Mol Ecol. 2007 Jun;16(11):2223-35
17561886
J Clin Microbiol. 2005 May;43(5):2236-40
15872248