Abstract
The classic definition of antimicrobial susceptibility to antifungal drugs ignores the persistence of subpopulations that survive in the presence of a drug. Even in entirely clonal populations, small subpopulations of yeast can grow in the presence of a drug, sometimes up to extremely high drug concentrations, such that they may be clinically relevant. Identifying and quantifying the incidence with which these subpopulations arise is an essential step in understanding how pathogenic yeast, such as Candida species (i.e., C. albicans, C. glabrata, C. auris, C. tropicalis, C. parapsilosis, and others) as well as Cryptococcus species, behave in response to antifungal therapeutics. Here we describe simple in vitro protocols for the quantification of drug responses with subpopulation resolution.
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References
Harsha MV, Venkatachalam S, Pooja M, Paranjothy M (2017) Emerging fungal pathogens – a major threat to human life. Int J Pharm Sci Res 8(5):1923–1934. https://doi.org/10.13040/IJPSR.0975-8232
Lamoth F, Lockhart SR, Berkow EL, Calandra T (2018) Changes in the epidemiological landscape of invasive candidiasis. J Antimicrob Chemother 73:i4. https://doi.org/10.1093/jac/dkx444
Sader HS, Pignatari AC (1994) E test: a novel technique for antimicrobial susceptibility testing. Sao Paulo Med J 112:635–638. https://doi.org/10.1590/s1516-31801994000400003
zur Wiesch PA, Kouyos R, Engelstädter J et al (2011) Population biological principles of drug-resistance evolution in infectious diseases. Lancet Infect Dis 11:236–247. https://doi.org/10.1016/S1473-3099(10)70264-4
Berman J, Krysan DJ (2020) Drug resistance and tolerance in fungi. Nat Rev Microbiol 18:319–331. https://doi.org/10.1038/s41579-019-0322-2
Rosenberg A, Ene IV, Bibi M et al (2018) Antifungal tolerance is a subpopulation effect distinct from resistance and is associated with persistent candidemia. Nat Commun 9:2470
Sionov E, Lee H, Chang YC, Kwon-Chung KJ (2010) Cryptococcus neoformans overcomes stress of azole drugs by formation of disomy in specific multiple chromosomes. PLoS Pathog 6(4):e1000848. https://doi.org/10.1371/journal.ppat.1000848
Sionov E, Chang YC, Kwon-Chung KJ (2013) Azole heteroresistance in cryptococcus neoformans: emergence of resistant clones with chromosomal disomy in the mouse brain during fluconazole treatment. Antimicrob Agents Chemother 57:5127–5130. https://doi.org/10.1128/AAC.00694-13
Stone NRH, Rhodes J, Fisher MC et al (2019) Dynamic ploidy changes drive fluconazole resistance in human cryptococcal meningitis. J Clin Invest 123:999. https://doi.org/10.1172/JCI124516
Gerstein AC, Rosenberg A, Hecht I, Berman J (2016) DiskimageR: quantification of resistance and tolerance to antimicrobial drugs using disk diffusion assays. Microbiol (United Kingdom) 162:1059–1068. https://doi.org/10.1099/mic.0.000295
White E (2019) ColonyScanalyser – an image analysis tool for measuring micro-organism colony growth. https://erik-white.github.io/ColonyScanalyser/
White E (2019) Mosey – multi scanner timed interval imaging tool for Windows https://github.com/Erik-White/Mosey
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Lyons, N., Berman, J. (2023). Protocols for Measuring Tolerant and Heteroresistant Drug Responses of Pathogenic Yeasts. In: Krysan, D.J., Moye-Rowley, W.S. (eds) Antifungal Drug Resistance. Methods in Molecular Biology, vol 2658. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3155-3_6
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DOI: https://doi.org/10.1007/978-1-0716-3155-3_6
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