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Inducing and Quantifying Clostridium difficile Spore Formation

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Book cover Clostridium difficile

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1476))

Abstract

The Gram-positive nosocomial pathogen Clostridium difficile induces sporulation during growth in the gastrointestinal tract. Sporulation is necessary for this obligate anaerobe to form metabolically dormant spores that can resist antibiotic treatment, survive exit from the mammalian host, and transmit C. difficile infections. In this chapter, we describe a method for inducing C. difficile sporulation in vitro. This method can be used to study sporulation and maximize spore purification yields for a number of C. difficile strain backgrounds. We also describe procedures for visualizing spore formation using phase-contrast microscopy and for quantifying the efficiency of sporulation using heat resistance as a measure of functional spore formation.

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Acknowledgments

K.F. is supported by T32 AI055402 from the National Institute of Allergy and Infectious Disease. A.S. is a Pew Scholar in the Biomedical Sciences, supported by The Pew Charitable Trusts; work in this manuscript was supported by start-up funds from Award Number P20 GM103496 and R01GM108684 from the National Institute of General Medical Sciences. The content is solely the responsibility of the authors and does not necessarily reflect the views of the Pew Charitable Trusts, the National Institute of Allergy and Infectious Disease, the National Institute of General Medical Sciences, or the National Institute of Health.

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Authors and Affiliations

  1. Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Drive, Burlington, VT, 05405, USA

    Aimee Shen, Kelly A. Fimlaid & Keyan Pishdadian

  2. Program in Cellular, Molecular & Biomedical Sciences, University of Vermont, Burlington, VT, 05405, USA

    Kelly A. Fimlaid

Authors
  1. Aimee Shen
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  2. Kelly A. Fimlaid
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  3. Keyan Pishdadian
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Corresponding author

Correspondence to Aimee Shen .

Editor information

Editors and Affiliations

  1. Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, United Kingdom

    Adam P. Roberts

  2. Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, United Kingdom

    Peter Mullany

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Shen, A., Fimlaid, K.A., Pishdadian, K. (2016). Inducing and Quantifying Clostridium difficile Spore Formation. In: Roberts, A., Mullany, P. (eds) Clostridium difficile. Methods in Molecular Biology, vol 1476. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6361-4_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6361-4_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6359-1

  • Online ISBN: 978-1-4939-6361-4

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