A flow cytometry method for assessing M. tuberculosis responses to antibiotics

Charlotte L. Hendon-Dunn, Stephen R. Thomas, Stephen C. Taylor, Joanna Bacon*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Citation (Scopus)

Abstract

Traditional drug susceptibility methods can take several days or weeks of incubation between drug exposure and confirmation of sensitivity or resistance. In addition, these methods do not capture information about viable organisms that are not immediately culturable after drug exposure. Here, we present a rapid fluorescence detection method for assessing the susceptibility of M. tuberculosis to antibiotics. Fluorescent markers Calcein violet-AM and SYTOX-green are used for measuring cell viability or cell death and to capture information about the susceptibility of the whole population and not just those bacteria that can grow in media postexposure. Postexposure to the antibiotic, the method gives a rapid readout of drug susceptibility, as well as insights into the concentration and time-dependent drug activity following antibiotic exposure.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages51-57
Number of pages7
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1736
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
Funding was received from Department of Health Grant in Aid and the National Institute of Health Research. The views expressed in this publication are those of the authors and not necessarily those of Public Health England, the National Institute for Health Research, or the Department of Health. The research leading to this method also received funding from the Innovative Medicines Initiative Joint Undertaking under grant agreement n°115337, resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007-2013) and EFPIA companies’ in-kind contribution. We acknowledge Professor Philip Marsh for his guidance and constructive input.

Publisher Copyright:
© 2018, Springer Science+Business Media, LLC.

Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.

Keywords

  • Antibiotic susceptibility
  • Calcein violet-AM
  • Flow cytometry
  • Mycobacterium tuberculosis
  • SYTOX-green

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