Restriction analysis of an amplified  rodA  gene fragment to distinguish  Aspergillus fumigatus var. ellipticus from  Aspergillus fumigatus var. fumigatus.
A previous multidisciplinary study indicated that gliotoxin-producing Aspergillus fumigatus Fresen. isolates from silage commodities mostly belonged to its variant A. fumigatus var. ellipticus Raper & Fennell. Sequence analysis revealed the presence of a single nucleotide polymorphism at five positions in a fragment of the rodA gene (coding for a hydrophobin rodletA protein) between Aspergillus fumigatus var. fumigatus and Aspergillus fumigatus var. ellipticus. A method was developed to distinguish these two types of isolates based on restriction analysis of this rodA gene fragment using the HinfI restriction enzyme. In addition, in silico analysis of 113 rodA gene fragments retrieved from GenBank was performed and confirmed the suitability of this method. In conclusion, the method developed in this study allows easy distinction between A. fumigatus var. fumigatus and its variant ellipticus. In combination with the earlier developed PCR-restriction fragment length polymorphism method of Staab et al. (2009, J Clin Microbiol 47: 2079), this method is part of a sequencing-independent identification scheme that allows for rapid distinction between similar species/variants within Aspergillus section Fumigati, specifically A. fumigatus, A. fumigatus var. ellipticus, Aspergillus lentulus Balajee & K.A. Marr, Neosartorya pseudofischeri S.W. Peterson and Neosartorya udagawae Y. Horie, Miyaji & Nishim.
Technology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Melle, Belgium. els.vanpamel@ilvo.vlaanderen.be
Els
E
Van Pamel
Els
E
Daeseleire
Nikki
N
De Clercq
Lieve
L
Herman
Annemieke
A
Verbeken
Marc
M
Heyndrickx
Geertrui
G
Vlaemynck
D001232:Aspergillus fumigatus / Q000145:classification* / Q000235:genetics*; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D016533:Mycological Typing Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D004271:DNA, Fungal; D005656:Fungal Proteins; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases