Validation of an expanded, in-house library and an optimized preparation method for the identification of fungal isolates using MALDI-TOF mass spectrometry

微生物学 背景(考古学) 曲霉 色谱法 红色毛癣菌 样品制备 基质辅助激光解吸/电离 生物 质谱法 化学 解吸 抗真菌 吸附 古生物学 有机化学
作者
Margarita Estreya Zvezdánova,Manuel González de Aledo,José Israel López-Mirones,Jesús Almeda,Andrés Canut,Carmen Castro,Carmen Cabrejas Gómez,Silvia Hernáez,Marina Oviaño,María Ercibengoa,Míriam Alkorta,Patricia Muñóz,David Rodríguez‐Temporal,Belén Rodríguez‐Sánchez
出处
期刊:Medical Mycology [Oxford University Press]
卷期号:61 (4) 被引量:3
标识
DOI:10.1093/mmy/myad038
摘要

Abstract The goal of this study was to validate an optimized sample preparation method for filamentous fungal isolates coupled with the use of an in-house library for the identification of moulds using Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) in a multicenter context. For that purpose, three Spanish microbiology laboratories participated in the identification of 97 fungal isolates using MALDI-TOF MS coupled with the Filamentous Fungi library 3.0 (Bruker Daltonics) and an in-house library containing 314 unique fungal references. The isolates analyzed belonged to 25 species from the genus Aspergillus, Fusarium, Scedosporium/Lomentospora, the Mucorales order and the Dermatophytes group. MALDI-TOF MS identification was carried out from hyphae resuspended in water and ethanol. After a high-speed centrifugation step, the supernatant was discarded and the pellet submitted to a standard protein extraction step. The protein extract was analyzed with the MBT Smart MALDI Biotyper system (Bruker Daltonics). The rate of accurate, species-level identification obtained ranged between 84.5% and 94.8% and the score values were 1.8 for 72.2–94.9% of the cases. Two laboratories failed to identify only one isolate of Syncephalastrum sp. and Trichophyton rubrum, respectively and three isolates could not be identified in the third center (F. proliferatum, n = 1; T.interdigitale, n = 2). In conclusion, the availability of an effective sample preparation method and an extended database allowed high rates of correct identification of fungal species using MALDI-TOF MS. Some species, such as Trichophyton spp. are still difficult to identify. Although further improvements are still required, the developed methodology allowed the reliable identification of most fungal species.
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