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首页> 外文期刊>Journal of Clinical Microbiology >Development of a Clinically Comprehensive Database and a Simple Procedure for Identification of Molds from Solid Media by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry
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Development of a Clinically Comprehensive Database and a Simple Procedure for Identification of Molds from Solid Media by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

机译：利用矩阵辅助激光解吸电离-飞行时间质谱从固体介质中鉴定霉菌的临床综合数据库和简单程序的开发

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Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a powerful tool for the rapid and highly accurate identification of clinical pathogens but has not been utilized extensively in clinical mycology due to challenges in developing an effective protein extraction method and the limited databases available. Here, we developed an alternate extraction procedure and constructed a highly stringent database comprising 294 individual isolates representing 76 genera and 152 species. To our knowledge, this is the most comprehensive clinically relevant mold database developed to date. When challenged with 421 blinded clinical isolates from our institution, by use of the BioTyper software, accurate species-level (score of ≥2.0) and genus-level (score of ≥1.7) identifications were obtained for 370 (88.9%) and 18 (4.3%) isolates, respectively. No isolates were misidentified. Of the 33 isolates (7.8%) for which there was no identification (score of <1.7), 25 were basidiomycetes not associated with clinical disease and 8 were Penicillium species that were not represented in the database. Our library clearly outperformed the manufacturer's database that was obtained with the instrument, which identified only 3 (0.7%) and 26 (6.2%) isolates at species and genus levels, respectively. Identification was not affected by different culture conditions. Implementation into our routine workflow has revolutionized our mycology laboratory efficiency, with improved accuracy and decreased time for mold identification, eliminating reliance on traditional phenotypic features.
机译：基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF MS）是快速，高度准确地鉴定临床病原体的有力工具，但由于开发有效的蛋白质提取方法面临挑战，因此尚未在临床真菌学中广泛使用方法和可用的有限数据库。在这里，我们开发了一种替代提取程序，并构建了一个高度严格的数据库，其中包含代表76个属和152个物种的294个分离株。据我们所知，这是迄今为止开发的最全面的临床相关模具数据库。当受到来自我们机构的421种盲目临床分离株的挑战时，通过使用BioTyper软件，获得了370种（88.9％）和18种（88.9％）的准确物种水平（≥2.0）和属水平（≥1.7）的鉴定。分别为4.3％）。没有隔离株被误认。在没有鉴定（得分<1.7）的33株分离株中（7.8％），25株与临床疾病无关的担子菌，8株在数据库中未显示的青霉菌。我们的文库明显胜过使用该仪器获得的制造商数据库，该数据库分别在物种和属水平上仅鉴定了3个（0.7％）和26（6.2％）分离株。鉴定不受不同培养条件的影响。常规工作流程的实施彻底改变了真菌学实验室的效率，提高了准确性，减少了模具鉴定的时间，从而消除了对传统表型特征的依赖。

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