Real8208;Time Viability Assay for Fungal cells

Ref ID: 18513

Author:

Paulo de Boer, Alie de Kat Angelino-Bart, Peter Punt, Remco Kort

Author address:

TNO, Microbiology & Systems Biology, P.O. Box 360, 3700 AJ, Zeist, The Netherlands

Full conference title:

11 th European Conference on Fungal Genetics

Abstract:

Detection of live microorganisms, including fungi, is currently performed by enumeration of colony forming units
(CFUs) on agar plates. Although reliable and widely accepted, this method has disadvantages. Determination of
CFUs is timely, laborious and limited to readily culturable microorganisms. Cultivation8208;independent methods are
available nowadays, but they mostly involve multiple incubation steps for assessment of the number of viable cells
and do not discriminate between dead or live microorganisms. Here a novel method is presented, capable of
specifically monitoring living fungal and yeast cells (and bacteria too) in a real8208;time manner.
The assay is based on a novel viability criterium, the ability of cells to maintain a neutral pH in an acidic
environment. Therefore, we searched for probes that show a fluorescent signal in a neutral environment, and not
in an acidic environment. Since fluorescence is only produced in cells with a neutral pH (i.e. living cells), the
fluorescent intensity is a measure for the amount of viable cells.
We have identified a number of probes that allow real8208;time viability (RTV) assays of fungal samples. These probes
have succesfully been tested on Aspergillus niger and A. fumigatus,   Saccharomyces cerevisiae, and Candida
albicans. Based on these results it is expected that the RTV assay will work for other fungal species too. The RTV
assay for fungi opens ways to assess in a fast, automated manner the viability in fungal samples such as spore
batches, anti8208;fungal treated fungal cells and all other applications where viability of fungal cells are of interest.

Abstract Number: PR8.8

Conference Year: 2012

Link to conference website: http://www.ecfg.info/images/Abstract_Book_Electronic.pdf

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