The clinical diagnosis of dermatophytosis and identification of dermatophytes face challenges due to reliance on culture-based methods. Rapid, cost-effective detection techniques for volatile organic compounds (VOCs) have been developed for other microorganisms, but their application to dermatophytes is limited. This study explores using VOCs as diagnostic markers for dermatophytes. We compared VOC profiles across different dermatophyte taxa using solid-phase microextraction (SPME) and advanced analytical methods: gas chromatography-mass spectrometry (GC-MS) and comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC-TOFMS). We analyzed 47 dermatophyte strains from 15 taxa grown on sheep wool, including clinically significant species. Additionally, we examined phylogenetic relationships among the strains to correlate genetic relatedness with metabolite production. Our results showed that GC×GC-TOFMS offered superior resolution but similar differentiation of VOC profiles compared to GC-MS. VOC spectra allowed reliable distinction of taxonomic units at the species level and below, however, these distinctions showed only a slight correlation with phylogenetic data. We identified pan-dermatophyte and species- or strain-specific VOC profiles, indicating their potential for rapid, non-invasive detection of dermatophyte infections, including epidemic strains. These patterns could enable future taxa-specific identification. Our study highlights the potential of VOCs as tools for dermatophyte taxonomy and diagnosis.

Department of Botany Faculty of Science Charles University Prague Czech Republic

Department of Genetics and Microbiology Faculty of Science Charles University Prague Czech Republic

Institute for Environmental Studies Faculty of Science Charles University Benátská 2 Prague 2 12801 Czech Republic

Laboratory of Environmental Biotechnology Institute of Microbiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Environmental Microbiology Institute of Microbiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Fungal Genetics and Metabolism Institute of Microbiology of the Czech Academy of Sciences Prague Czech Republic

Organic and Biological Analytical Chemistry Group Chemistry Department University of Liège Liège Belgium

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