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利用手持式质谱探头直接从培养物和临床样本中鉴定临床相关细菌。

Identifying Clinically Relevant Bacteria Directly from Culture and Clinical Samples with a Handheld Mass Spectrometry Probe.

机构信息

Department of Chemistry, The University of Texas at Austin, Austin, TX 78712, USA.

Department of Pediatrics, Division of Pediatric Infectious Diseases, J.W. Riley Hospital for Children, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

出版信息

Clin Chem. 2022 Nov 3;68(11):1459-1470. doi: 10.1093/clinchem/hvac147.

Abstract

BACKGROUND

Rapid identification of bacteria is critical to prevent antimicrobial resistance and ensure positive patient outcomes. We have developed the MasSpec Pen, a handheld mass spectrometry-based device that enables rapid analysis of biological samples. Here, we evaluated the MasSpec Pen for identification of bacteria from culture and clinical samples.

METHODS

A total of 247 molecular profiles were obtained from 43 well-characterized strains of 8 bacteria species that are clinically relevant to osteoarticular infections, including Staphylococcus aureus, Group A and B Streptococcus, and Kingella kingae, using the MasSpec Pen coupled to a high-resolution mass spectrometer. The molecular profiles were used to generate statistical classifiers based on metabolites that were predictive of Gram stain category, genus, and species. Then, we directly analyzed samples from 4 patients, including surgical specimens and clinical isolates, and used the classifiers to predict the etiologic agent.

RESULTS

High accuracies were achieved for all levels of classification with a mean accuracy of 93.3% considering training and validation sets. Several biomolecules were detected at varied abundances between classes, many of which were selected as predictive features in the classifiers including glycerophospholipids and quorum-sensing molecules. The classifiers also enabled correct identification of Gram stain type and genus of the etiologic agent from 3 surgical specimens and all classification levels for clinical specimen isolates.

CONCLUSIONS

The MasSpec Pen enables identification of several bacteria at different taxonomic levels in seconds from cultured samples and has potential for culture-independent identification of bacteria directly from clinical samples based on the detection of metabolic species.

摘要

背景

快速鉴定细菌对于防止抗菌药物耐药和确保患者良好转归至关重要。我们开发了 MasSpec Pen,这是一种基于手持式质谱的设备,可实现对生物样本的快速分析。在此,我们评估了 MasSpec Pen 对培养和临床样本中细菌的鉴定能力。

方法

我们使用 MasSpec Pen 结合高分辨率质谱仪,对 8 种与骨关节炎感染相关的临床重要细菌(金黄色葡萄球菌、A 组和 B 组链球菌以及金氏金氏菌)的 43 种特征明确的菌株进行了总共 247 个分子谱的检测。使用这些分子谱,基于可预测革兰氏染色类别、属和种的代谢物,生成统计分类器。然后,我们直接分析了 4 名患者的样本,包括手术标本和临床分离株,并使用分类器预测病原体。

结果

考虑到训练集和验证集,所有分类水平的准确率均很高,平均准确率为 93.3%。在不同类别之间检测到多种生物分子,其丰度存在差异,其中许多被选为分类器中的预测特征,包括甘油磷脂和群体感应分子。这些分类器还能够正确识别 3 个手术标本中的病原体的革兰氏染色类型和属,以及临床标本分离株的所有分类水平。

结论

MasSpec Pen 能够在数秒内从培养样本中鉴定出不同分类水平的多种细菌,并且有可能通过检测代谢物种,直接从临床样本中进行无需培养的细菌鉴定。

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