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基质辅助激光解吸/电离飞行时间质谱法用于快速鉴定真菌性鼻-鼻窦炎病原体

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid identification of fungal rhinosinusitis pathogens.

作者信息

Huang Yanfei, Wang Jinglin, Zhang Mingxin, Zhu Min, Wang Mei, Sun Yufeng, Gu Haitong, Cao Jingjing, Li Xue, Zhang Shaoya, Lu Xinxin

机构信息

Department of Laboratory Medicine, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China.

State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing, PR China.

出版信息

J Med Microbiol. 2017 Mar;66(3):328-333. doi: 10.1099/jmm.0.000435. Epub 2017 Mar 20.

Abstract

PURPOSE

Filamentous fungi are among the most important pathogens, causing fungal rhinosinusitis (FRS). Current laboratory diagnosis of FRS pathogens mainly relies on phenotypic identification by culture and microscopic examination, which is time consuming and expertise dependent. Although matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS has been employed to identify various fungi, its efficacy in the identification of FRS fungi is less clear.

METHODOLOGY

A total of 153 FRS isolates obtained from patients were analysed at the Clinical Laboratory at the Beijing Tongren Hospital affiliated to the Capital Medical University, between January 2014 and December 2015. They were identified by traditional phenotypic methods and Bruker MALDI-TOF MS (Bruker, Biotyper version 3.1), respectively. Discrepancies between the two methods were further validated by sequencing.

RESULTS

Among the 153 isolates, 151 had correct species identification using MALDI-TOF MS (Bruker, Biot 3.1, score ≥2.0 or 2.3). MALDI-TOF MS enabled identification of some very closely related species that were indistinguishable by conventional phenotypic methods, including 1/10 Aspergillus versicolor, 3/20 Aspergillus flavus, 2/30 Aspergillus fumigatus and 1/20 Aspergillus terreus, which were misidentified by conventional phenotypic methods as Aspergillus nidulans, Aspergillus oryzae, Aspergillus japonicus and Aspergillus nidulans, respectively. In addition, 2/2 Rhizopus oryzae and 1/1 Rhizopus stolonifer that were identified only to the genus level by the phenotypic method were correctly identified by MALDI-TOF MS.

CONCLUSION

MALDI-TOF MS is a rapid and accurate technique, and could replace the conventional phenotypic method for routine identification of FRS fungi in clinical microbiology laboratories.

摘要

目的

丝状真菌是最重要的病原体之一,可引起真菌性鼻-鼻窦炎(FRS)。目前FRS病原体的实验室诊断主要依靠培养和显微镜检查进行表型鉴定,这既耗时又依赖专业知识。尽管基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱已被用于鉴定各种真菌,但其在鉴定FRS真菌方面的功效尚不太明确。

方法

2014年1月至2015年12月期间,首都医科大学附属北京同仁医院临床实验室对从患者身上获得的153株FRS分离株进行了分析。它们分别通过传统表型方法和布鲁克MALDI-TOF质谱(布鲁克,Biotyper 3.1版)进行鉴定。两种方法之间的差异通过测序进一步验证。

结果

在153株分离株中,151株使用MALDI-TOF质谱(布鲁克,Biot 3.1,分数≥2.0或2.3)进行了正确的物种鉴定。MALDI-TOF质谱能够鉴定一些通过传统表型方法无法区分的非常相近的物种,包括1/10的杂色曲霉、3/20的黄曲霉、2/30的烟曲霉和1/20的土曲霉,这些在传统表型方法中分别被错误鉴定为构巢曲霉、米曲霉、日本曲霉和构巢曲霉。此外,通过表型方法仅鉴定到属水平的2/2米根霉和1/1匍枝根霉通过MALDI-TOF质谱被正确鉴定。

结论

MALDI-TOF质谱是一种快速准确的技术,可在临床微生物实验室中替代传统表型方法用于FRS真菌的常规鉴定。

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