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Master Thesis

Détection de mutations associées à la résistance aux antifongiques chez Candida spp. par pyroséquençage haut débit

Abstract : Azole and echinocandin antifungals are increasingly used for the prevention and the treatment of invasive candidiasis, leading to the development of resistant strains possibly associated with clinical failure. This study aimed at detecting mutations involved in antifungal resistance in Candida spp. using a next-generation sequencing (NGS) technology: high-troughput pyrosequencing. Several genes from a single isolate, several isolates and several Candida species were simultaneously and reliably sequenced using the GS Junior® platform (Roche Diagnostics). The experimental strategy allowed the detection of mutations associated with echinocandin resistance. Several unreported mutations were detected by the analysis of genes involved in azole resistance. Sequential isolates obtained from patients colonized or infected by C. albicans, C. glabrata or C. parapsilosis before and after antifungal treatment were sequenced. Neither appearance nor disappearance of any mutation involved in antifungal resistance could have been detected. Sequencing of supplemental genes and isolates will provide the detection of new mutations and a better comprehension of resistance mechanisms in Candida spp. This technology could also be applied to the analysis of antifungal resistance in other fungal pathogens including Aspergillus spp. Finally, the use of NGS technologies in clinical practice could allow the early detection of isolates prone to the development of phenotypic resistance or clinical failure.
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Master Thesis
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https://dumas.ccsd.cnrs.fr/dumas-01254639
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Submitted on : Tuesday, January 12, 2016 - 3:10:11 PM
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  • HAL Id : dumas-01254639, version 1

Citation

Cécile Garnaud. Détection de mutations associées à la résistance aux antifongiques chez Candida spp. par pyroséquençage haut débit. Sciences pharmaceutiques. 2013. ⟨dumas-01254639⟩

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