Abstract

Online MALDI-TOF mass spectrometry applications, such as MSI-2, have been shown to help identify dermatophytes, but recurrent errors are still observed between phylogenetically close species. The objective of this study was to assess different approaches to reduce the occurrence of such errors by adding new reference spectra to the MSI-2 application. Nine libraries were set up, comprising an increasing number of spectra obtained from reference strains that were submitted to various culture durations on two distinct culture media: Sabouraud gentamicin chloramphenicol medium and IDFP Conidia medium. The final library included spectra from 111 strains of 20 species obtained from cultures on both media collected every three days after the appearance of the colony. The performance of each library was then analyzed using a cross-validation approach. The spectra acquisitions were carried out using a Microflex Bruker spectrometer. Diversifying the references and adding spectra from various culture media and culture durations improved identification performance. The percentage of correct identification at the species level rose from 63.4 to 91.7% when combining all approaches. Nevertheless, residual confusion between close species, such as Trichophyton rubrum, Trichophyton violaceum and Trichophyton soudanense, remained. To distinguish between these species, mass spectrometry identification should take into account basic morphological and/or clinico-epidemiological features.

Highlights

  • Dermatophytes are a monophyletic clade of keratinophilic fungi (Arthrodermataceae) that are frequent skin pathogens in human and animals [1,2]

  • The strains were reseeded on two culture plates: Sabouraud medium with gentamicin and chloramphenicol but without cycloheximide (Oxoid, Dardilly, France) (SGC) and ID-fungi plate (IDFP) medium

  • T. rubrum complex, T. interdigitale/T. mentagrophytes, T. equinum/T. tonsurans and Microsporum spp., the results differed according to the complex

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Summary

Introduction

Dermatophytes are a monophyletic clade of keratinophilic fungi (Arthrodermataceae) that are frequent skin pathogens in human and animals [1,2]. Until the 2000 s, dermatophyte identification was mostly based on macroscopic and microscopic morphological features. This conventional approach requires well-trained professionals and extended culture time. MALDI-TOF mass spectrometry appeared as an alternative allowing earlier and standardized identification of fungal colonies. Many studies have been published since 2008 [3,4]. Demonstrating the interest of this identification method. The best performances have been obtained with “in-house” libraries and not commercial libraries [5]. Recurrent errors still occurred regarding closely related species such as T. rubrum, T. violaceum and T

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