A MALDI-TOF MS procedure for clinical dermatophyte species identification in the routine laboratory
A MALDI-TOF MS procedure for clinical dermatophyte species identification in the routine laboratory
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DOI：
10.3109/13693786.2013.781691
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发表时间：
2013-10-01
期刊：
影响因子：
2.9
通讯作者：
Ranque, Stephane
中科院分区：
文献类型：
--
作者：
L'Ollivier, Coralie;Cassagne, Carole;Ranque, Stephane
皮肤真菌的常规鉴定需要很长的周转时间和高度熟练的真菌学家。我们最近开发了一种标准化的基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）分析，以常规识别具有潜在临床意义的霉菌。本研究的目的是确定这种相同的检测方法是否也可以用于在常规实验室环境中识别临床皮肤真菌。包括放线菌酮在培养基和孵育时间的影响进行了测试后，建立了一个参考光谱库，其中包括48个良好的特点分离的17种皮肤癣菌。然后使用该文库对这些相同的分离株进行前瞻性鉴定。无论是否存在放线菌酮或孵育时间如何，基于MALDI-TOF MS的鉴定都是有效的，因为130/133（97.8%）的临床分离株得到了适当鉴定。两个犬小孢子菌分离株产生了无信息的光谱和一个M。audouinii分离物被错误鉴定。由于MALDI-TOF MS分析只需要一个小菌落，因此可以在3-6天内获得准确的鉴定，特别是在其特征形态特征出现之前。因此，识别周转时间显着减少相比，传统的形态识别所需的。总之，这种标准化的基于MALDI-TOF MS的丝状真菌鉴定程序有效地鉴定了临床皮肤真菌分离株，并大大减少了常规临床实验室的响应时间。
The conventional identification of dermatophytes requires a long turnaround time and highly skilled mycologists. We have recently developed a standardized matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) assay to routinely identify molds of potential clinical significance. This study objective was to determine if this same assay could also be employed to identify clinical dermatophytes in the routine laboratory setting. The effects of the inclusion of cycloheximide in the culture medium and incubation time were tested after building a reference spectra library that included 48 well-characterized isolates of 17 dermatophyte species. Then these same isolates were prospectively identified using this library. MALDI-TOF MS-based identification was effective regardless of the presence of cycloheximide or incubation time as 130/133 (97.8%) of the clinical isolates were appropriately identified. Two Microsporum canis isolates yielded uninformative spectra and one M. audouinii isolate was misidentified. Since one only requires a small colony for MALDI-TOF MS analysis, accurate identifications were obtained in 3-6 days and, specifically, before the appearance of their characteristic morphological features. Consequently, identification turnaround time was dramatically reduced as compared to that needed for conventional morphological identification. In conclusion, this standardized MALDI-TOF MS-based identification procedure for filamentous fungi effectively identifies clinical dermatophyte isolates and drastically reduces the response times in the routine clinical laboratory.