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1 Department of Medical Microbiology, University College London, London W1T 4JF, UK
2 Imperial College London, Centre for Microbiology and Infection, Flowers Building, Armstrong Rd, London SW7 2AZ, UK
3 Medical Microbiology and Infectious Diseases, Location Lukas, Gelre hospitals, Albert Schweitzerlaan 31, 7300 DS Apeldoorn, The Netherlands
Correspondence
Helen D. Donoghue
h.donoghue{at}ucl.ac.uk
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.
Abbreviations: MTB, Mycobacterium tuberculosis
Present address: Molecular Immunology (Hill Group), Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK.
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