Abstract
Monitoring persister cells can be extremely difficult due to their transient and stochastic nature, their low abundance, and their resemblance to Viable But Non-Culturable Cells (VBNCs). To date, the predominant method consists of determining the survival rate of a bacterial population after antibiotic treatment as a function of time or antibiotic concentration. Unfortunately, this method is limited, as it shows high levels of dispersion of the data around the mean, making interpretation difficult. Furthermore, additional reproducibility problems arise from the lack of a standard method, different research groups using different protocols. Here, we describe a standard and optimized method for monitoring E. coli persister cells at the population level allowing for maximal reproducibility.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K (2004) Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 230(1):13–18
Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S (2004) Bacterial persistence as a phenotypic switch. Science 305(5690):1622–1625. doi:10.1126/science.1099390
Joers A, Kaldalu N, Tenson T (2010) The frequency of persisters in Escherichia coli reflects the kinetics of awakening from dormancy. J Bacteriol 192(13):3379–3384. doi:10.1128/JB.00056-10
Orman MA, Brynildsen MP (2013) Establishment of a method to rapidly assay bacterial persister metabolism. Antimicrob Agents Chemother 57(9):4398–4409. doi:10.1128/AAC.00372-13
Korch SB, Henderson TA, Hill TM (2003) Characterization of the hipA7 allele of Escherichia coli and evidence that high persistence is governed by (p)ppGpp synthesis. Mol Microbiol 50(4):1199–1213
Hansen S, Lewis K, Vulic M (2008) Role of global regulators and nucleotide metabolism in antibiotic tolerance in Escherichia coli. Antimicrob Agents Chemother 52(8):2718–2726. doi:10.1128/AAC.00144-08
Sezonov G, Joseleau-Petit D, D’Ari R (2007) Escherichia coli physiology in Luria-Bertani broth. J Bacteriol 189(23):8746–8749. doi:10.1128/JB.01368-07
Neidhardt FC, Bloch PL, Smith DF (1974) Culture medium for enterobacteria. J Bacteriol 119(3):736–747
Blattner FR, Plunkett G 3rd, Bloch CA, Perna NT, Burland V, Riley M, Collado-Vides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y (1997) The complete genome sequence of Escherichia coli K-12. Science 277(5331):1453–1462
Luidalepp H, Joers A, Kaldalu N, Tenson T (2011) Age of inoculum strongly influences persister frequency and can mask effects of mutations implicated in altered persistence. J Bacteriol 193(14):3598–3605. doi:10.1128/JB.00085-11
Maisonneuve E, Shakespeare LJ, Jorgensen MG, Gerdes K (2011) Bacterial persistence by RNA endonucleases. Proc Natl Acad Sci U S A 108(32):13206–13211. doi:10.1073/pnas.1100186108
Acknowledgement
Research in Van Melderen’s lab was funded by FNRS (FRSM 3.4621.12), the Interuniversity Attraction Poles Programme initiated by the Belgian Science Policy Office (MICRODEV), the Fonds Jean Brachet and the Fondation David and Alice Van Buuren. F.G. was supported by the FRIA.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Goormaghtigh, F., Van Melderen, L. (2016). Optimized Method for Measuring Persistence in Escherichia coli with Improved Reproducibility. In: Michiels, J., Fauvart, M. (eds) Bacterial Persistence. Methods in Molecular Biology, vol 1333. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2854-5_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2854-5_4
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2853-8
Online ISBN: 978-1-4939-2854-5
eBook Packages: Springer Protocols