Rapid Detection and Simultaneous Strain Differentiation of Mycobacterium tuberculosis Complex Bacteria by Spoligotyping

  • Henri O. F. Molhuizen
  • Annelies E. Bunschoten
  • Leo M. Schouls
  • Jan D. A. van Embden
Part of the Methods in Molecular Biology™ book series (MIMB, volume 101)


The discovery of polymorphic DNA in Mycobacterium tuberculosis has led to several methods to differentiate climcal isolates. The most widely used method is based on the mobility of the insertion element IS6110, which is present in virtually all M tuberculosis isolates, usually in multiple copies. IS6110-based fingerprinting has been extremely useful for epidemiological studies such as outbreak investigations, assessment of risk factors for recent transmissions, and for studying the dissemination of multiple drug resistant strains (1,2).


  1. 1.
    Small, P. M, Embden, J D. M, and Embden, J D A van (1994) Molecular epidemiology of tuberculosis, in Tuberculosis: Pathogenesis, Protection and Control (Bloom, B., ed) ASM Press, Washington, DC, 569–582.Google Scholar
  2. 2.
    Small, P. M., Hopewell, P. C., Singh, S. P., Paz, A, Parsonnet, J, Ruston, D C., Schechter, G F., Daley, C. L, and Schoolnik, G. K (1994) The epidemiology of tuberculosis in San Francisco. N. Eng J. Med 330, 1703–1708CrossRefGoogle Scholar
  3. 3.
    Groenen, P. M. A., Bunschoten, A. E., Soolingen, D. van, and Embden, J. D. A. van (1993) Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; Application for strain differentiation by a novel method. Mol Microbiol 10(5), 1057–1065PubMedCrossRefGoogle Scholar
  4. 4.
    Haas, W. H., Butler,: W R., Woodley, C H, and Crawford, J T. (1993) Mixed-linker polymerase chain reaction a new method for rapid fingerprinting of isolates of the Mycobacterium tuberculosis complex. J Clin. Microbiol 31, 1293–1298PubMedGoogle Scholar
  5. 5.
    Palittapongarnpim, P., Chomic, S, Fanning, A., and Kunimoto, D (1993) DNA fingerprinting of Mycobacterium tuberculosis by ligation-mediated polymerase chain reaction Nucl Acids Res 3, 761,762CrossRefGoogle Scholar
  6. 6.
    Palittapongarnpim, P, Chomic, S, Fanning, A., and Kunimoto, D (1993) DNA fragment length polymorphism analysis of Mycobacterium tuberculosis isolates by arbitrarily primed polymerasee chain reaction. J. Inf Dis 167, 975–978.Google Scholar
  7. 7.
    Ross, B C and Dwyer, B. (1993) Rapid, simple method for typing isolates of Mycobacterium tuberculosis by using the polymerase chain reaction. J Clin Microbiol 31, 329–334.PubMedGoogle Scholar
  8. 8.
    Hermans, P. W. M., Soolingen, D., van Bik, E. M., Haas, P. E. W., de, Dale, J. W., and Embden, J. D. A. van (1991) The insertion element IS987 from Mycobacterium bovis BCG is located in a hot spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect. Immunol 59, 2695–2705Google Scholar
  9. 9.
    Kaufhold, A., Podbielski, A., Baumgarten, G., Blokpoel, M., Top, J., and Schouls L. (1994) Rapid typing of group A streptococci by the use of DNA amplification and nonradioactive allele specific oligonucleotide probes FEMS Microbiol Lett 119, 19–26PubMedCrossRefGoogle Scholar
  10. 10.
    Kamerbeek, J, Schouls, L, Agterveld, M., van, Kolk, A, van Agterveld, D, van Soolingen, D., Kuijper, J. E, Bunschoten, A E, Molhuizen,: M, Shaw, R, Goyal, M., and van Embden, J. (1996) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol 35, 907–914Google Scholar
  11. 11.
    Soolingen, D., van, Qian, L., Haas, P. E. W., D van, Douglas, J. T., Traore, H, Portaels, F., Qing, H Z, Enksaikan, D., Nymadawa, P., and Embden, J. D. A, van (1995) Predominance of a single genotype of Mycobacterium tuberculosis in countries of East Asia. J Clin Microbiol 33, 3234–3238.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1998

Authors and Affiliations

  • Henri O. F. Molhuizen
  • Annelies E. Bunschoten
  • Leo M. Schouls
  • Jan D. A. van Embden

There are no affiliations available

Personalised recommendations