Semi-automated repetitive sequence-based PCR amplification for species of the Scedosporium apiospermum complex.
OBJECTIVE
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 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.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).
EA 3800, Université de Rouen, 76031 Rouen, France Laboratoire de Parasitologie-Mycologie, CHU Charles-Nicolle, 76031 Rouen, France.
Olivier
O
Matray
L'UNAM Université, Université d'Angers, Groupe d'Etude des Interactions Hôte-Pathogène, EA 3142, 49933 Angers, France Department of Biology, Faculty of Sciences, University Moulay Ismail, Meknes, Morocco.
Abdelmounaim
A
Mouhajir
L'UNAM Université, Université d'Angers, Groupe d'Etude des Interactions Hôte-Pathogène, EA 3142, 49933 Angers, France.
Sandrine
S
Giraud
L'UNAM Université, Université d'Angers, Groupe d'Etude des Interactions Hôte-Pathogène, EA 3142, 49933 Angers, France.
Charlotte
C
Godon
EA 3800, Université de Rouen, 76031 Rouen, France.
Gilles
G
Gargala
EA 3800, Université de Rouen, 76031 Rouen, France.
Franck
F
Labbé
L'UNAM Université, Université d'Angers, Groupe d'Etude des Interactions Hôte-Pathogène, EA 3142, 49933 Angers, France Centre Hospitalier Universitaire, Laboratoire de Parasitologie-Mycologie, Institut de Biologie en Santé-PBH, 49933 Angers, France.
Amandine
A
Rougeron
EA 3800, Université de Rouen, 76031 Rouen, France.
Jean-Jacques
JJ
Ballet
Department of Biology, Faculty of Sciences, University Moulay Ismail, Meknes, Morocco.
Rachid
R
Zouhair
L'UNAM Université, Université d'Angers, Groupe d'Etude des Interactions Hôte-Pathogène, EA 3142, 49933 Angers, France Centre Hospitalier Universitaire, Laboratoire de Parasitologie-Mycologie, Institut de Biologie en Santé-PBH, 49933 Angers, France.
Jean-Philippe
JP
Bouchara
EA 3800, Université de Rouen, 76031 Rouen, France Laboratoire de Parasitologie-Mycologie, CHU Charles-Nicolle, 76031 Rouen, France loic.favennec@chu-rouen.fr.
Loïc
L
Favennec
D003550:Cystic Fibrosis / Q000382:microbiology; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D009181:Mycoses / Q000382:microbiology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D021681:Scedosporium / Q000145:classification / Q000235:genetics*; D013183:Sputum / Q000382:microbiology
Cystic fibrosis; Genotyping; Scedosporium apiospermum species complex; Species identification; rep-PCR