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Insertion Element IS6110-Based Restriction Fragment Length Polymorphism Genotyping of Mycobacterium tuberculosis

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Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 465))

Abstract

DNA fingerprinting techniques are based on genome variation and form the basis of molecular epidemiology studies of tuberculosis. A number of markers are in use for the molecular differentiation of Mycobacterium tuberculosis isolates by DNA fingerprinting. One of these markers is the IS6110 insertion element, which may be present in up to 25 copies per M. tuberculosis genome. Variation in both the number and location of the IS6110 elements makes it a very useful marker of strain genotype. IS6110-based DNA fingerprinting is globally considered as the reference genotyping technique for M. tuberculosis isolates. This method is based on visualization of restriction fragment length polymorphisms using a labeled probe derived from IS6110. In this chapter, the method of IS6110 DNA fingerprinting is explained in such a way that it can be easily duplicated by molecular epidemiologists and will give reproducible results.

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Acknowledgments

We would like to thank Ms. Talita Lotz for providing the IS6110-based RFLP image.

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Authors and Affiliations

  1. DST/NRF Centre of Excellence in Biomedical Tuberculosis Research / MRC Centre for Molecular and Cellular Biology, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Health Sciences, Stellenbosch University, PO Box 19063, Tygerberg, South Africa, 7505

    Robin M. Warren

  2. Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and The London, Queen Mary’s School of Medicine and Dentistry, London, UK

    Paul D. van Helden PhD & Nicolaas C. Gey van Pittius PhD

Authors
  1. Robin M. Warren
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  2. Paul D. van Helden PhD
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  3. Nicolaas C. Gey van Pittius PhD
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Corresponding author

Correspondence to Robin M. Warren .

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Editors and Affiliations

  1. Queen Mary's School of Med. & Dentistry, Inst. Cell & Molecular Science, Barts and the London, Newark Street 4, London, E1 2AT, United Kingdom

    Tanya Parish

  2. Queen Mary's School of Med. & Dentistry, Inst. Cell & Molecular Science, Barts and the London, Newark Street 4, London, E1 2AT, United Kingdom

    Amanda Claire Brown

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Warren, R.M., van Helden, P.D., van Pittius, N.C.G. (2009). Insertion Element IS6110-Based Restriction Fragment Length Polymorphism Genotyping of Mycobacterium tuberculosis . In: Parish, T., Brown, A. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 465. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-207-6_24

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  • DOI: https://doi.org/10.1007/978-1-59745-207-6_24

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-58829-889-8

  • Online ISBN: 978-1-59745-207-6

  • eBook Packages: Springer Protocols

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