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Master Thesis

Évaluation d'une technique de rep-PCR semi-automatisée pour le génotypage de souches du complexe Pseudallescheria boydii / Scedosporium apiospermum au cours de la mucoviscidose

Abstract : Species of the Scedosporium apiospermum complex rank the second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF). It is clearly established now that these fungi may disseminate in case of immunodeficiency and that a chronic colonization of the airways by these pathogens may hinder the success of lung transplantation. In this study, we develop a new genotyping method to investigate the epidemiology of the airway colonization by these fungi. 63 multiple and sequential isolates of S. apiospermum collected from 9 CF patients, and selected among those previously studied by random amplification of polymorphic DNA (RAPD) by Defontaine et at. (2002) were analyzed. using the automated typing system DiversiLab® (bioMerieux) based on PCR amplification of repetitive sequences. Two different rep-PCR kits were compared; the DiverSiLab® Aspergillus and the DiversiLab® Fungal kit. The amplification products were separated by capillary electrophoresison; Each obtained profiles were then compared using. the DiversiLab® software, In addition, species identification of these isolates was clarified by sequencing TUB locus of the P-tubulin gene. The Fungal kit appeared to be more discriminating than the Aspergillus kit. Additionally, rep-PCR usually confirmed the colonization patterns described by RAPD. Only one patients showed distinct genotypes; In conclusion, the automated typing system DiversiLab® proved to be an easy and efficient Method to investigate the molecular epidemiology of the airway colonization by P. boydii / S. apiospermum complex species in CF. Moreover, our results confirm the capacity of the different species from this complex to chronically colonize the airways of CF patients despite the repeated antifungal cures.
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Master Thesis
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https://dumas.ccsd.cnrs.fr/dumas-00916347
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Submitted on : Tuesday, December 10, 2013 - 10:39:14 AM
Last modification on : Monday, December 2, 2019 - 10:24:01 AM
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  • HAL Id : dumas-00916347, version 1

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Julien Matray. Évaluation d'une technique de rep-PCR semi-automatisée pour le génotypage de souches du complexe Pseudallescheria boydii / Scedosporium apiospermum au cours de la mucoviscidose. Sciences pharmaceutiques. 2013. ⟨dumas-00916347⟩

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