Viable quantitative PCR for assessing the response of Candida albicans to antifungal treatment.
Appl Microbiol Biotechnol
; 97(1): 341-9, 2013 Jan.
Article
em En
| MEDLINE
| ID: mdl-23132341
ABSTRACT
Propidium monoazide (PMA) or ethidium bromide monoazide (EMA) treatment has been used before nucleic acid detection methods, such as PCR, to distinguish between live and dead cells using membrane integrity as viability criterion. The performance of these DNA intercalating dyes was compared in many studies utilizing different microorganisms. These studies demonstrated that EMA and PMA differ in their abilities to identify nonviable cells from mixed cell populations, depending on the microorganism and the nature of the sample. Due to this heterogeneity, both dyes were used in the present study to specifically distinguish dead from live Candida albicans cells using viable quantitative PCR (qPCR). The viable qPCR was optimized, and the best results were obtained when pre-treating the cells for 10 min in the dark with 25 µM EMA followed by continuous photoactivation for 15 min. The suitability of this technique to distinguish clotrimazole- and fluconazole-treated C. albicans cells from untreated cells was then assessed. Furthermore, the antifungal properties of two commercial essential oils (Thymus vulgaris and Matricaria chamomilla) were evaluated. The viable qPCR method was determined to be a feasible technique for assessing the viability of C. albicans after drug treatment and may help to provide a rapid diagnostic and susceptibility testing method for fungal infections, especially for patients treated with antifungal therapies.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Candida albicans
/
Contagem de Colônia Microbiana
/
Viabilidade Microbiana
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Reação em Cadeia da Polimerase em Tempo Real
/
Antifúngicos
Tipo de estudo:
Evaluation_studies
/
Prognostic_studies
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article