Matray, O. and Mouhajir, A. and Giraud, S. and Godon, C. and Gargala, G. and Labbé, F. and Rougeron, A. and Ballet, J.-J. and Zouhair, R. and Bouchara, J.-P. and Favennec, L. (2016) Semi-automated repetitive sequence-based PCR amplification for species of the Scedosporium apiospermum complex. Medical Mycology, 54 (4). pp. 1-11.

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Purpose: The Scedosporium apiospermum species complex usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), but little is known about the molecular epidemiology of the airway colonization. Methods: Polymerase chain reaction (PCR) amplification of repetitive sequences (rep- PCR) was applied to the retrospective analysis of a panel of isolates already studied by random amplification of polymorphic DNA (RAPD) and comprising 63 isolates recovered from sputa from 9 CF patients. Results were compared to those obtained previously by RAPD, and herein by beta-tubulin (TUB) gene sequencing and Multilocus Sequence Typing (MLST). Results: Within the panel of isolates studied, S. apiospermum sensu stricto and Scedosporium boydii, as expected, were the predominant species with 21 and 36 isolates, respectively. Four isolates from one patient were identified as Scedosporium aurantiacum, whereas two isolates belonged to the Pseudallescheria ellipsoidea subgroup of S. boydii. rep-PCR analysis of these isolates clearly differentiated the three species and P. ellipsoidea isolates, whatever the rep-PCR kit used, and also permitted strain differentiation. When using the mold primer kit, results from rep-PCR were in close agreement with those obtained by MLST. For both S. apiospermum and S. boydii, 8 genotypes were differentiated by rep-PCR andMLST compared to 10 by RAPD. All S. aurantiacum isolates shared the same RAPD genotype and exhibited the same rep-PCR profile and sequence type. Conclusions: These results illustrate the efficacy of rep-PCR for both species identification within the S. apiospermum complex and genotyping for the two major species of this complex. Abstract presentation: Part of this work was presented during the 18th Congress of the International Society for Human and Animal Mycology, Berlin (Germany), June 2012. S. Giraud, C. Godon, A. Rougeron, J.P. Bouchara and L. Favennec are members of the ECMM/ISHAM working group on Fungal respiratory infections in Cystic Fibrosis (Fri-CF). © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.

Item Type: Article
Uncontrolled Keywords: beta tubulin; fungal DNA, clinical article; Conference Paper; cystic fibrosis; fungal strain; fungus isolation; gene amplification; gene sequence; genotype; human; multilocus sequence typing; nonhuman; nucleotide sequence; polymerase chain reaction; Pseudallescheria; Pseudallescheria ellipsoidea; random amplified polymorphic DNA; repetitive sequence based polymerase chain reaction amplification; Scedosporium apiospermum; Scedosporium apiospermum complex; Scedosporium apiospermum sensu stricto; Scedosporium aurantiacum; Scedosporium boydii; Scedosporium minutisporum; species identification; strain difference; classification; genetics; microbiology; molecular typing; mycosis; phylogeny; polymerase chain reaction; procedures; Scedosporium; sputum, Cystic Fibrosis; Humans; Molecular Typing; Mycoses; Phylogeny; Polymerase Chain Reaction; Random Amplified Polymorphic DNA Technique; Scedosporium; Sputum
Subjects: Medicine
Depositing User: Administrateur Eprints Administrateur Eprints
Last Modified: 31 Jan 2020 15:49

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