Rapid identification of 6328 isolates of pathogenic yeasts using MALDI-ToF MS and a simplified, rapid extraction procedure that is compatible with the Bruker Biotyper platform and database
Rapid identification of 6328 isolates of pathogenic yeasts using MALDI-ToF MS and a simplified, rapid extraction procedure that is compatible with the Bruker Biotyper platform and database
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DOI：
10.1093/mmy/myv085
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发表时间：
2016-01-01
期刊：
影响因子：
2.9
通讯作者：
Johnson, Elizabeth M.
中科院分区：
文献类型：
--
作者：
Fraser, Mark;Brown, Zoe;Johnson, Elizabeth M.
从临床样本中快速准确地鉴定酵母菌分离株是必不可少的，因为它们天生具有可变的抗真菌敏感性特征，并且提出了物种特异性抗真菌敏感性解释断点。在这里，我们评估了MALDI-ToF质谱分析用于鉴定临床分离的致病酵母的效用。我们实验室开发了一种简化、快速的提取方法，应用于包括71种不同酵母菌的6343株分离物，然后使用Bruker Microflex进行MALDI-ToF质谱分析，并使用提供的Bruker数据库评估所得光谱。MALDI-ToF ms共鉴定出6328/6343株（99.8%）分离株。简化的提取方案可正确鉴定93.6%的分离株，无需费力的全提取，全提取后可鉴定出394株（6.2%）分离株。两种提取方法的临床相关鉴定都是使用提供的Bruker数据库实现的，不需要生成定制的内部数据库，这些数据库是使用适应的提取方法获得的配置文件创建的。事实上，使用我们的方法获得的平均LogScores与使用推荐的、已发布的完整提取过程获得的LogScores一样健壮。然而，内部数据库可以为不寻常或很少遇到的生物体提供有用的额外识别工具。最后，所提出的方法允许从我们实验室提交的初始培养中直接正确鉴定超过75%的分离株，而无需在标准化真菌学培养基上进行额外的继代培养。
Rapid and accurate identification of yeast isolates from clinical samples is essential, given their innately variable antifungal susceptibility profiles, and the proposal of species-specific antifungal susceptibility interpretive breakpoints. Here we have evaluated the utility of MALDI-ToF MS analysis for the identification of clinical isolates of pathogenic yeasts. A simplified, rapid extraction method, developed in our laboratory, was applied to 6343 isolates encompassing 71 different yeast species, which were then subjected to MALDI-ToF MS analysis using a Bruker Microflex and the resulting spectra were assessed using the supplied Bruker database. In total, 6328/6343 (99.8%) of isolates were correctly identified by MALDI-ToF MS. Our simplified extraction protocol allowed the correct identification of 93.6% of isolates, without the need for laborious full extraction, and a further 394 (6.2%) of isolates could be identified after full extraction. Clinically relevant identifications with both extraction methods were achieved using the supplied Bruker database and did not require the generation of bespoke, in-house databases created using profiles obtained with the adapted extraction method. In fact, the mean LogScores obtained using our method were as robust as those obtained using the recommended, published full extraction procedures. However, an in-house database can provide a useful additional identification tool for unusual or rarely encountered organisms. Finally, the proposed methodology allowed the correct identification of over 75% of isolates directly from the initial cultures referred to our laboratory, without the requirement for additional sub-culture on standardised mycological media.