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Semi-automated repetitive sequence-based PCR amplification for species of the Scedosporium apiospermum complex

Abstract : 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 Sce-dosporium boydii, as expected, were the predominant species with 21 and 36 isolates, respectively. Four isolates from one patient were identified as Scedosporium auranti-acum, 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 and MLST 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
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Olivier Matray, Abdelmounaim Mouhajir, Sandrine Giraud, Charlotte Godon, Gilles Gargala, et al.. Semi-automated repetitive sequence-based PCR amplification for species of the Scedosporium apiospermum complex. Medical Mycology, Oxford University Press, 2016, 54 (4), pp.409-419. ⟨10.1093/mmy/myv080⟩. ⟨hal-02317900⟩



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