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利用基质辅助激光解吸电离飞行时间质谱技术进行真菌鉴定。

Dermatophyte identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry.

机构信息

Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Hilton 868, 200 First Street SW, Rochester, MN 55905, USA.

出版信息

J Clin Microbiol. 2011 Dec;49(12):4067-71. doi: 10.1128/JCM.01280-11. Epub 2011 Sep 28.

DOI:10.1128/JCM.01280-11
PMID:21956979
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3232958/
Abstract

The performance of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometer (MS) for the identification of dermatophytes from clinical cultures was compared to that of dermatophyte identification using 28S rRNA gene sequencing. The MALDI Biotyper library (MBL; version 3.0) was used alone and in combination with a supplemented library containing an additional 20 dermatophyte spectra (S-MBL). Acquired spectra were interpreted using both the manufacturer-recommended scores (genus, ≥1.7; species, ≥2.0) and adjusted cutoff values established by this study (genus, ≥1.5; species, ≥1.7); identifications required a minimum 10% difference in scores between the top two different organisms to be considered correct. One hundred well-characterized, archived dermatophyte isolates and 71 fresh dermatophyte cultures were evaluated using both libraries and both sets of cutoff criteria. Collectively, the S-MBL significantly outperformed the MBL at both the genus (93% versus 37.4%; P < 0,0001) and species (59.6% versus 20.5%; P < 0.0001) levels when using the adjusted score criteria. Importantly, application of the lowered cutoff values significantly improved genus (P = 0.005)- and species (P < 0.0001)-level identification for the S-MBL, without leading to an increase in misidentifications. MALDI-TOF MS is a cost-effective and rapid alternative to traditional or molecular methods for dermatophyte identification, provided that the reference library is supplemented to sufficiently encompass clinically relevant, intraspecies strain diversity.

摘要

本研究比较了 Bruker Biotyper 基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱仪(MS)与 28S rRNA 基因测序用于鉴定临床培养物中皮肤癣菌的性能。单独使用 MALDI Biotyper 库(MBL;版本 3.0)和包含另外 20 种皮肤癣菌光谱的补充库(S-MBL)进行鉴定。使用制造商推荐的评分(属,≥1.7;种,≥2.0)和本研究建立的调整后的临界值(属,≥1.5;种,≥1.7)来解释获得的光谱;鉴定需要两个不同生物体之间的得分差异最小为 10%,才能被认为是正确的。使用这两个库和两套临界值标准,对 100 个经过充分特征描述的存档皮肤癣菌分离株和 71 个新鲜皮肤癣菌培养物进行了评估。在使用调整后的评分标准时,S-MBL 在属(93%比 37.4%;P < 0.0001)和种(59.6%比 20.5%;P < 0.0001)水平上的性能明显优于 MBL。重要的是,应用降低的临界值显著提高了 S-MBL 的属(P = 0.005)和种(P < 0.0001)水平的鉴定,而不会导致错误鉴定增加。MALDI-TOF MS 是一种具有成本效益且快速的传统或分子方法替代方法,用于皮肤癣菌鉴定,前提是参考库得到补充,以充分涵盖临床相关的种内菌株多样性。

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