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High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium

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CRISPR

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1311))

Abstract

Spoligotyping was developed almost 18 years ago and still remains a popular first-lane genotyping technique to identify and subtype Mycobacterium tuberculosis complex (MTC) clinical isolates at a phylogeographic level. For other pathogens, such as Salmonella enterica, recent studies suggest that specifically designed spoligotyping techniques could be interesting for public health purposes. Spoligotyping was in its original format a reverse line-blot hybridization method using capture probes designed on “spacers” and attached to a membrane’s surface and a PCR product obtained from clustered regularly interspaced short palindromic repeats (CRISPRs). Cowan et al. and Fabre et al. were the first to propose a high-throughput Spoligotyping method based on microbeads for MTC and S. enterica serotype Typhimurium, respectively. The main advantages of the high-throughput Spoligotyping techniques we describe here are their low cost, their robustness, and the existence (at least for MTC) of very large databases that allow comparisons between spoligotypes from anywhere.

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Acknowledgments

The authors would like to thank Jian Zhang, Marie Accou-Demartin, Lucile Sontag, Saïana de Romans, Catherine Lim and Laëtitia Fabre. M. François Topin and M. Jan van Gils from Luminex BV, The Netherlands, are also acknowledged for technical support.

Author information

Authors and Affiliations

  1. Microbiology Department, Institut de Biologie Intégrative de la Cellule (I2BC), CEA, CNRS, Université Paris-Sud, Rue Gregor Mendel, Building 400, Room 205-208, F-91405, Orsay-Cedex, France

    Christophe Sola

  2. Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela

    Edgar Abadia

  3. Institut Pasteur, Unité des Bactéries Pathogènes Entériques, Paris, France

    Simon Le Hello & François-Xavier Weill

Authors
  1. Christophe Sola
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  2. Edgar Abadia
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  3. Simon Le Hello
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  4. François-Xavier Weill
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Corresponding author

Correspondence to Christophe Sola .

Editor information

Editors and Affiliations

  1. Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden

    Magnus Lundgren

  2. Department of Molecular Biology MIMS, Umeå University, Umeå, Sweden

    Emmanuelle Charpentier

  3. Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand

    Peter C. Fineran

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Sola, C., Abadia, E., Le Hello, S., Weill, FX. (2015). High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium. In: Lundgren, M., Charpentier, E., Fineran, P. (eds) CRISPR. Methods in Molecular Biology, vol 1311. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2687-9_6

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  • DOI: https://doi.org/10.1007/978-1-4939-2687-9_6

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2686-2

  • Online ISBN: 978-1-4939-2687-9

  • eBook Packages: Springer Protocols

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