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基质辅助激光解吸电离飞行时间质谱法用于正血培养物中细菌鉴定的声捕获。

Acoustic trapping for bacteria identification in positive blood cultures with MALDI-TOF MS.

机构信息

Department of Biomedical Engineering, Lund University , 221 00 Lund, Sweden.

出版信息

Anal Chem. 2014 Nov 4;86(21):10560-7. doi: 10.1021/ac502020f. Epub 2014 Oct 16.

Abstract

Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is currently changing the clinical routine for identification of microbial pathogens. One important application is the rapid identification of bacteria for the diagnosis of bloodstream infections (BSI). A novel approach based on acoustic trapping and an integrated selective enrichment target (ISET) microchip that improves the sample preparation step for this type of analysis is presented. The method is evaluated on clinically relevant samples in the form of Escherichia coli infected blood cultures. It is shown that noncontact acoustic trapping enables capture, enrichment, and washing of bacteria directly from the complex background of crude blood cultures. The technology replaces centrifugation-based separation with a faster and highly automated sample preparation method that minimizes manual handling of hazardous pathogens. The presented method includes a solid phase extraction step that was optimized for enrichment of the bacterial proteins and peptides that are used for bacterial identification. The acoustic trapping-based assay provided correct identification in 12 out 12 cases of E. coli positive blood cultures with an average score of 2.19 ± 0.09 compared to 1.98 ± 0.08 when using the standard assay. This new technology opens up the possibility to automate and speed up an important and widely used diagnostic assay for bloodstream infections.

摘要

基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)正在改变微生物病原体鉴定的临床常规。一个重要的应用是快速鉴定细菌,用于诊断血流感染(BSI)。本文提出了一种基于声捕获和集成选择性富集靶(ISET)微芯片的新方法,该方法改进了此类分析的样品制备步骤。该方法以大肠杆菌感染的血培养物的临床相关样本进行评估。结果表明,非接触式声捕获能够直接从复杂的粗血培养物背景中捕获、富集和洗涤细菌。该技术用更快、高度自动化的样品制备方法替代了基于离心的分离,最大限度地减少了对危险病原体的人工处理。所提出的方法包括优化用于富集细菌蛋白和肽的固相萃取步骤,这些蛋白和肽用于细菌鉴定。基于声捕获的检测方法在 12 例大肠杆菌阳性血培养物的检测中得到了正确的鉴定,平均得分为 2.19±0.09,而使用标准检测方法的平均得分为 1.98±0.08。这项新技术为自动化和加速血流感染的一种重要且广泛使用的诊断检测方法提供了可能。

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