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基质辅助激光解吸电离飞行时间质谱法鉴定丝状真菌:三种不同样品制备方法的评估及内部物种截断值的验证

Identification of Filamentous Fungi by MALDI-TOF Mass Spectrometry: Evaluation of Three Different Sample Preparation Methods and Validation of an In-House Species Cutoff.

作者信息

Honsig Claudia, Selitsch Brigitte, Hollenstein Marlene, Vossen Matthias G, Spettel Kathrin, Willinger Birgit

机构信息

Division of Clinical Microbiology, Department of Laboratory Medicine, Medical University of Vienna, 1090 Vienna, Austria.

Department of Laboratory Medicine, Medical University of Vienna, 1090 Vienna, Austria.

出版信息

J Fungi (Basel). 2022 Apr 10;8(4):383. doi: 10.3390/jof8040383.

DOI:10.3390/jof8040383
PMID:35448614
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9025689/
Abstract

Invasive infections caused by filamentous fungi constitute a leading cause of morbidity and mortality in immunocompromised patients. Rapid and reliable identification of filamentous fungi is essential for the early initiation of appropriate treatment. In the present study, 230 filamentous fungi isolates identified by conventional methods were investigated using MALDI-TOF MS (Bruker Daltonics, Bremen, Germany) in combination with the Filamentous Fungi Library 3.0 provided by the manufacturer. Three different sample preparation methods were applied as recommended by the manufacturer and identification rates were compared using the criteria provided by the manufacturer. Application of the more time-consuming sample preparation methods clearly improved identification at the species level. Thus, the identification rate increased from 48.9% using the simplest method to 76.1% with the most laborious procedure. Misidentifications did not occur. Furthermore, the reliability of an in-house threshold for species identification was investigated. The reduced threshold increased the rate of isolates correctly identified at the species level by up to 86.4%. As no misidentification was made at the genus level and only one misidentification of minor significance occurred at the species level, this threshold could be validated for routine use in our laboratory. In conclusion, regarding the high identification rates achieved, this commercial platform proved suitable for implementation in routine diagnosis.

摘要

丝状真菌引起的侵袭性感染是免疫功能低下患者发病和死亡的主要原因。快速、可靠地鉴定丝状真菌对于尽早开始适当治疗至关重要。在本研究中,使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS,德国不来梅布鲁克道尔顿公司)结合制造商提供的丝状真菌库3.0,对通过传统方法鉴定的230株丝状真菌分离株进行了研究。按照制造商的建议应用了三种不同的样品制备方法,并使用制造商提供的标准比较了鉴定率。应用耗时较长的样品制备方法明显提高了物种水平的鉴定率。因此,鉴定率从使用最简单方法时的48.9%提高到最繁琐程序时的76.1%。未出现错误鉴定。此外,还研究了用于物种鉴定的内部阈值的可靠性。降低阈值使物种水平正确鉴定的分离株率提高了86.4%。由于在属水平未出现错误鉴定,且在物种水平仅出现一例意义不大的错误鉴定,该阈值可在我们实验室用于常规验证。总之,鉴于所取得的高鉴定率,该商业平台被证明适用于常规诊断。

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