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基质辅助激光解吸电离飞行时间质谱法鉴定毛霉目真菌

Identification of Mucorales by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry.

作者信息

Schwarz Patrick, Guedouar Houssem, Laouiti Farah, Grenouillet Frédéric, Dannaoui Eric

机构信息

Department of Internal Medicine, Respiratory and Critical Care Medicine, University Hospital Marburg, D-35043 Marburg, Germany.

Center for Invasive Mycoses and Antifungals, Philipps University Marburg, D-35037 Marburg, Germany.

出版信息

J Fungi (Basel). 2019 Jul 2;5(3):56. doi: 10.3390/jof5030056.

Abstract

More than 20 different species of Mucorales can be responsible for human mucormycosis. Accurate identification to the species level is important. The morphological identification of Mucorales is not reliable, and the currently recommended identification standard is the molecular technique of sequencing the internal transcribed spacer regions. Nevertheless, matrix-assisted laser desorption ionization time-of-flight mass spectrometry has been shown to be an accurate alternative for the identification of bacteria, yeasts, and even filamentous fungi. Therefore, 38 Mucorales isolates, belonging to 12 different species or varieties, mainly from international collections, including 10 type or neo-type strains previously identified by molecular methods, were used to evaluate the usefulness of matrix-assisted laser desorption ionization time-of-flight mass spectrometry for the identification of human pathogenic Mucorales to the species level. One to three reference strains for each species were used to create a database of main spectrum profiles, and the remaining isolates were used as test isolates. A minimum of 10 spectra was used to build the main spectrum profile of each database strain. Interspecies discrimination for all the isolates, including species belonging to the same genus, was possible. Twenty isolates belonging to five species were used to test the database accuracy, and were correctly identified to the species level with a log-score >2. In summary, matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a reliable and rapid method for the identification of most of the human pathogenic Mucorales to the species level.

摘要

20多种不同的毛霉目真菌可导致人类毛霉病。准确鉴定到种水平很重要。毛霉目的形态学鉴定不可靠,目前推荐的鉴定标准是对内部转录间隔区进行测序的分子技术。然而,基质辅助激光解吸电离飞行时间质谱已被证明是鉴定细菌、酵母甚至丝状真菌的一种准确替代方法。因此,本研究使用了38株毛霉目分离株,分属于12个不同的种或变种,主要来自国际菌种保藏中心,其中包括10株先前通过分子方法鉴定的模式株或新模式株,以评估基质辅助激光解吸电离飞行时间质谱在将人类致病毛霉目真菌鉴定到种水平方面的实用性。每个种使用1至3株参考菌株创建主要质谱图谱数据库,其余分离株用作测试菌株。每个数据库菌株至少使用10个光谱来构建主要质谱图谱。对所有分离株,包括同属的种,都能够进行种间鉴别。使用属于5个种的20株分离株测试数据库的准确性,这些菌株均以对数得分>2被正确鉴定到种水平。总之,基质辅助激光解吸电离飞行时间质谱是一种可靠且快速的方法,可将大多数人类致病毛霉目真菌鉴定到种水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56e3/6787577/8655fcefdb07/jof-05-00056-g001.jpg

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