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利用 MALDI-TOF 质谱技术鉴定丝状真菌,包括皮肤癣菌。

Identification of filamentous fungi including dermatophytes using MALDI-TOF mass spectrometry.

机构信息

Department of Microbiology, University Hospital, Olomouc, Czech Republic.

Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

出版信息

Folia Microbiol (Praha). 2022 Feb;67(1):55-61. doi: 10.1007/s12223-021-00917-6. Epub 2021 Sep 9.

Abstract

Identification of filamentous fungi based on morphological features is the most available approach used in clinical mycology laboratories. However, MALDI-TOF mass spectrometry is currently invaluable for identification of microorganisms because of its rapidity, simplicity, and accuracy. This study aimed to find the optimal way of identifying filamentous fungi using MALDI-TOF MS.The sample comprised 193 isolates of filamentous fungi. The identification started with morphological assessment. Then isolates were identified using MALDI-TOF MS, both directly from culture and following culture in liquid media with extraction. Subsequently, identification of 20 selected isolates was compared by sequencing of the benA gene, ITS1-5,8-ITS2, and D1-D2 LSU regions.Based on morphological criteria, 17 genera of fungi were identified. With MALDI-TOF MS performed directly from culture, nine isolates were identified to the genus level and 184 to the species level, with a total of 75 species being noted. With the MALDI-TOF MS extraction method, 190 isolates were identified to the species level, with 43 species being noted. The rates of agreement between identification using morphology and the MALDI-TOF MS direct method were 58.55% at the genus level and 22.24% at the species level. The rates of agreement between identification using morphology and the MALDI-TOF MS extraction method were 84.97% at the genus level and 46.11% at the species level. Using sequencing, 87.5% agreement was found for identification with the MALDI-TOF MS extraction method, as compared with only 43.75% for the direct method.The results suggest that the optimal approach to identification of filamentous fungi is a combination of morphological features and MALDI-TOF MS using the extraction method.

摘要

基于形态特征的丝状真菌鉴定是临床真菌学实验室最常用的方法。然而,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)因其快速、简单和准确而对微生物的鉴定具有不可估量的价值。本研究旨在寻找使用 MALDI-TOF MS 鉴定丝状真菌的最佳方法。

该样本包括 193 株丝状真菌分离株。鉴定工作首先从形态评估开始。然后,通过 MALDI-TOF MS 直接从培养物以及在液体培养基中培养并提取后进行鉴定。随后,通过测序 benA 基因、ITS1-5.8-ITS2 和 D1-D2 LSU 区比较了 20 株选定分离株的鉴定结果。

根据形态学标准,鉴定出 17 个属的真菌。通过 MALDI-TOF MS 直接从培养物进行鉴定,有 9 个分离株鉴定到属水平,184 个分离株鉴定到种水平,共鉴定出 75 个种。通过 MALDI-TOF MS 提取方法,有 190 个分离株鉴定到种水平,共鉴定出 43 个种。形态学鉴定与 MALDI-TOF MS 直接法鉴定的一致性在属水平为 58.55%,在种水平为 22.24%。形态学鉴定与 MALDI-TOF MS 提取法鉴定的一致性在属水平为 84.97%,在种水平为 46.11%。通过测序,与直接法相比,MALDI-TOF MS 提取法的鉴定结果一致性为 87.5%。

结果表明,丝状真菌鉴定的最佳方法是将形态特征与 MALDI-TOF MS 提取法相结合。

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