Evaluation of internal transcribed spacer 2-RFLP analysis for the identification of dermatophytes.
A total of 95 isolates, belonging to 33 species of five dermatophyte genera, i.e. Arthroderma (15 species), Chrysosporium (two), Epidermophyton (one), Microsporum (three) and Trichophyton (12), were studied using internal transcribed spacer 2 (ITS2)-PCR-RFLP analysis (ITS2-RFLP), consisting of amplification of the ITS2 region, restriction digestion with BstUI (CG/CG) and restriction fragment length determination by capillary electrophoresis. ITS2-RFLP analysis proved to be most useful for identification of species of the genera Arthroderma, Chrysosporium and Epidermophyton, but could not distinguish between several Trichophyton species. The identification results are in agreement with established and recent taxonomical insights into the dermatophytes; for example, highly related species also had closely related and sometimes difficult-to-discriminate ITS2-RFLP patterns. In some cases, several ITS2-RFLP groups could be distinguished within species, again mostly in agreement with the taxonomic delineations of subspecies and/or genomovars, confirming the relevance of ITS2-RFLP analysis as an identification technique and as a useful taxonomic approach.
Laboratory for Bacteriology Research, University of Ghent, B-9000 Ghent, Belgium.
Thierry
T
De Baere
CBS Fungal Diversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Richard
R
Summerbell
CBS Fungal Diversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Bart
B
Theelen
Division of Acute Medicine and Infectious Diseases, University Medical Centre Utrecht, Utrecht, The Netherlands.|CBS Fungal Diversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Teun
T
Boekhout
Laboratory for Bacteriology Research, University of Ghent, B-9000 Ghent, Belgium.
Mario
M
Vaneechoutte
D000818:Animals; D003883:Arthrodermataceae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D008969:Molecular Sequence Data; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer