We have shown that dermatophyte species can be easily identified on the basis of a DNA sequence encoding a part of the large-subunit (LSU) rRNA (28S rRNA) by using the MicroSeq D2 LSU rRNA Fungal Sequencing Kit. Two taxa causing distinct dermatophytoses were clearly distinguished among isolates of the Trichophyton mentagrophytes species complex.Dermatophytes are the main cause of superficial mycoses (9,15,16). These fungi have the capacity to invade keratinized tissue of humans or animals to produce infections that are generally restricted to the corneocytes of the skin, hair, and nails. Among the approximately 10 species isolated in Europe, Trichophyton rubrum and T. mentagrophytes are the most commonly observed, with frequencies varying from 27 to 74% and from 17 to 41%, respectively (13).Dermatophytes are usually identified on the basis of macroscopic appearance, together with microscopic examination of cultures. Important characteristics are the rate of growth, the shape and texture of the culture on solid media, color, diffusion of pigments into agar, and sporulation. However, identification of dermatophytes often remains difficult or uncertain because there are variations from one isolate to another. Recent advances in molecular biology and progress in technology have allowed the development of new techniques for species determination and strain typing in microbiology. The molecular approach used to identify fungi is often based on sequence analysis of the ribosomal DNA (rDNA) and in particular on the internal transcribed spacer (ITS). The polymorphism of the ITS1 and ITS2 regions flanking the DNA sequence encoding the 5.8S rRNA was previously shown to be suitable for the identification of clinically important yeasts (1), Aspergillus sp. (8),and dermatophyte species (3-5). In contrast, the gene coding for the small-subunit rRNA (18S rRNA) did not discriminate sufficiently between dermatophyte species (7).The MicroSeq D2 large-subunit (LSU) rRNA Fungal Sequencing Kit (Applied Biosystems, Rotkreuz, Switzerland) was recently developed to identify fungal species after amplification of a partial sequence of the DNA encoding the LSU rRNA (28S rDNA). The sequence of a given fungus can then be compared for identification with the rDNA sequences of the MicroSeq D2 Fungal database, which includes more than 500 validated sequences from different fungal species but not from dermatophytes. In the present study, we tested the MicroSeq D2 LSU rRNA Fungal Sequencing Kit by using it to identify dermatophyte species from patients referred to the mycological laboratory of the Department of Dermatology at the University Hospital in Lausanne, Switzerland (Table 1). Neotypes of different species and other reference strains (Table 2) were used for comparison of DNA sequences. This study allows the extension of the MicroSeq D2 Fungal database to the determination of dermatophytes.Isolates. Skin and nail scrapings and hair fragments were collected from patients with suspected mycoses. Routinely, one part of each sample was examine...