Abstract
We describe a rapid and simple in vitro method for development of Candida biofilms under static growth conditions. This 96-well microtiter-based method measures metabolic activity of sessile cells and can also be easily adapted for antifungal susceptibility testing. The entire procedure takes 2–3 days to complete, reliably quantifies biofilms, and provides reproducible results that are imperative toward the standardization of antifungal susceptibility testing of biofilms.
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References
Donlan RM (2002) Biofilms: microbial life on surfaces. Emerg Infect Dis 8(9):881–890
Nobile CJ, Johnson AD (2015) Candida albicans biofilms and human disease. Annu Rev Microbiol 69:71–92
Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284(5418):1318–1322
Uppuluri P, Lin L, Alqarihi A, Luo G, Youssef EG, Alkhazraji S et al (2018) The Hyr1 protein from the fungus Candida albicans is a cross kingdom immunotherapeutic target for Acinetobacter bacterial infection. PLoS Pathog 14(5):e1007056
Uppuluri P, Chaturvedi AK, Srinivasan A, Banerjee M, Ramasubramaniam AK, Kohler JR et al (2010) Dispersion as an important step in the Candida albicans biofilm developmental cycle. PLoS Pathog 6(3):e1000828
Ramage G, Saville SP, Thomas DP, Lopez-Ribot JL (2005) Candida biofilms: an update. Eukaryot Cell 4(4):633–638
Pierce CG, Uppuluri P, Tristan AR, Wormley FL Jr, Mowat E, Ramage G et al (2008) A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 3(9):1494–1500
Bujdakova H, Kulkova N, Cernakova L (2012) Susceptibility to caspofungin and fluconazole and Als1/Als3 gene expression in biofilm and dispersal cells of Candida albicans. Arh Hig Rada Toksikol 63(4):497–503
Pierce CG, Uppuluri P, Tummala S, Lopez-Ribot JL (2010) A 96 well microtiter plate-based method for monitoring formation and antifungal susceptibility testing of Candida albicans biofilms. J Vis Exp (44):2287
Uppuluri P, Dinakaran H, Thomas DP, Chaturvedi AK, Lopez-Ribot JL (2009) Characteristics of Candida albicans biofilms grown in a synthetic urine medium. J Clin Microbiol 47(12):4078–4083
Corte L, Casagrande Pierantoni D, Tascini C, Roscini L, Cardinali G (2019) Biofilm specific activity: a measure to quantify microbial biofilm. Microorganisms 7(3):73
Mamouei Z, Alqarihi A, Singh S, Xu S, Mansour MK, Ibrahim AS et al (2018) Alexidine dihydrochloride has broad spectrum activities against diverse fungal pathogens. mSphere 3(6):e00539-18
Pierce CG, Saville SP, Lopez-Ribot JL (2014) High-content phenotypic screenings to identify inhibitors of Candida albicans biofilm formation and filamentation. Pathog Dis 70(3):423–431
Singh S, Uppuluri P, Mamouei Z, Alqarihi A, Elhassan H, French S et al (2019) The NDV-3A vaccine protects mice from multidrug resistant Candida auris infection. PLoS Pathog 15(8):e1007460
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Uppuluri, P. (2022). A Simple 96-Well Plate-Based Method for Development of Candida Biofilms Under Static Conditions. In: Calderone, R. (eds) Candida Species. Methods in Molecular Biology, vol 2542. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2549-1_16
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DOI: https://doi.org/10.1007/978-1-0716-2549-1_16
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