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首页> 外文期刊>Journal of Clinical Microbiology >Comparison of the Bruker Biotyper and Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Mycobacteria Using Simplified Protein Extraction Protocols
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Comparison of the Bruker Biotyper and Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Mycobacteria Using Simplified Protein Extraction Protocols

机译:布鲁克Biotyper和Vitek MS基质辅助激光解吸电离的飞行时间质谱系统比较,用于使用简化的蛋白质提取方案鉴定分枝杆菌

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摘要

Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has recently been described as a fast and inexpensive method for the identification of mycobacteria. Although mycobacteria require extraction prior to MALDI-TOF MS analysis, previously published protocols have been relatively complex, involving significant hands-on time and materials not often found in the clinical laboratory. In this study, we tested two simplified protein extraction protocols developed at the University of Washington (UW) and by bioMérieux (BMX) for use with two different mass spectrometry platforms (the Bruker MALDI Biotyper and the bioMérieux Vitek MS, respectively). Both extraction protocols included vortexing with silica beads in the presence of ethanol. The commercial Bruker database was also augmented with an in-house database composed of 123 clinical Mycobacterium strains. A total of 198 clinical strains, representing 18 Mycobacterium species, were correctly identified to the species level 94.9% of the time when extracted using the UW protocol and compared to the augmented database. The BMX protocol and Vitek MS system resulted in correct species-level identifications for 94.4% of these strains. In contrast, only 79.3% of the strains were identified to the species level by the nonaugmented Bruker database, although the use of a lower identification score threshold (≥1.7) increased the identification rate to 93.9%, with two misidentifications that were unlikely to be clinically relevant. The two simplified protein extraction protocols described in this study are easy to use for identifying commonly encountered Mycobacterium species.
机译:基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)最近已被描述为鉴定分枝杆菌的一种快速而廉价的方法。尽管分枝杆菌需要在MALDI-TOF MS分析之前进行提取,但以前发表的方案相对复杂,涉及大量的动手时间和临床实验室中不常发现的材料。在这项研究中,我们测试了由华盛顿大学(UW)和bioMérieux(BMX)开发的两种简化的蛋白质提取方案,分别用于两种不同的质谱分析平台(分别为Bruker MALDI Biotyper和bioMérieuxVitek MS)。两种提取方案均包括在乙醇存在下用二氧化硅珠涡旋。商业化的布鲁克数据库也增加了由123种临床分枝杆菌菌株组成的内部数据库。使用UW方案提取并代表增强数据库时,代表18种分枝杆菌属的198株临床菌株在94.9%的时间内被正确鉴定到物种水平。 BMX协议和Vitek MS系统对94.4%的这些菌株进行了正确的物种级鉴定。相比之下,通过不增强的布鲁克数据库,只有79.3%的菌株被鉴定到物种水平,尽管使用较低的鉴定分数阈值(≥1.7)将鉴定率提高到93.9%,其中有两个错误鉴定不太可能是临床相关。本研究中描述的两种简化的蛋白质提取方案易于识别常见的分枝杆菌种类。

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