Institute of Microbiology, via Mirasole 22A, 6501 Bellinzona, Switzerland.
Med Mycol. 2013 Jul;51(5):514-21. doi: 10.3109/13693786.2012.746476. Epub 2012 Dec 11.
In this study we evaluated the suitability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of dermatophytes in diagnostic laboratories. First, a spectral database was built with 108 reference strains belonging to 18 species of the anamorphic genera Epidermophyton, Microsporum and Trichophyton. All strains were well characterized by morphological criteria and ITS sequencing (gold standard). The dendrogram resulting from MALDI-TOF mass spectra was almost identical with the phylogenetic tree based on ITS sequencing. Subsequently, MALDI-TOF MS SuperSpectra were created for the identification of Epidermophyton floccosum, Microsporium audouinii, M. canis, M. gypseum (teleomorph: Arthroderma gypseum), M. gypseum (teleomorph: A. incurvatum), M. persicolor, A. benhamiae (Tax. Entity 3 and Am-Eur. race), T. erinacei, T. interdigitale (anthropophilic and zoophilic populations), T. rubrum/T. violaceum, T. tonsurans and T. terrestre. Because T. rubrum and T. violaceum did not present enough mismatches, a SuperSpectrum covering both species was created, and differentiation between them was done by comparison of eight specific peptide masses. In the second part of this study, MALDI-TOF MS with the newly created SuperSpectra was tested using 141 clinical isolates representing nine species. Analyses were done with 3-day-old cultures. Results were compared to morphological identification and ITS sequencing; 135/141 (95.8%) strains were correctly identified by MALDI-TOF MS compared to 128/141 (90.8%) by morphology. Therefore, MALDI-TOF MS has proven to be a useful and rapid identification method for dermatophytes.
在这项研究中,我们评估了基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)在诊断实验室中鉴定皮肤癣菌的适用性。首先,我们构建了一个包含 108 株参考菌株的光谱数据库,这些菌株属于形态学属 Epidermophyton、Microsporum 和 Trichophyton 的 18 个种。所有菌株均通过形态学标准和 ITS 测序(金标准)得到了很好的鉴定。MALDI-TOF 质谱产生的系统发育树几乎与基于 ITS 测序的系统发育树相同。随后,我们为鉴定 Epidermophyton floccosum、Microsporium audouinii、M. canis、M. gypseum(无性型:Arthroderma gypseum)、M. gypseum(无性型:A. incurvatum)、M. persicolor、A. benhamiae(分类实体 3 和欧美种)、T. erinacei、T. interdigitale(亲兽型和兽源型)、T. rubrum/T. violaceum、T. tonsurans 和 T. terrestre 创建了 MALDI-TOF MS SuperSpectra。由于 T. rubrum 和 T. violaceum 没有足够的不匹配,因此创建了一个涵盖这两个种的 SuperSpectrum,并通过比较 8 种特定肽质量来区分它们。在本研究的第二部分,我们使用代表 9 个种的 141 株临床分离株对新创建的 SuperSpectra 进行了 MALDI-TOF MS 测试。分析采用 3 日龄培养物进行。将结果与形态学鉴定和 ITS 测序进行比较;与形态学鉴定相比,MALDI-TOF MS 正确鉴定了 135/141(95.8%)株,而正确鉴定了 128/141(90.8%)株。因此,MALDI-TOF MS 已被证明是一种有用且快速的皮肤癣菌鉴定方法。