MS applications in microbiology have increased significantly in the past 10 years, due in part to the proliferation of regulator‐approved commercial MALDI MS platforms for rapid identification of clinical infections. In parallel, with the expansion of MS technologies in the “omics” fields, novel MS‐based research efforts to characterize organismal as well as environmental microbiomes have emerged. Successful characterization of microorganisms found in complex mixtures of other organisms remains a major challenge for researchers and clinicians alike. Here, we review recent MS advances toward addressing that challenge. These include sample preparation methods and protocols, and established, for example, MALDI, as well as newer, for example, atmospheric pressure ionization (API) techniques. MALDI mass spectra of intact cells contain predominantly information on the highly expressed house‐keeping proteins used as biomarkers. The API methods are applicable for small biomolecule analysis, for example, phospholipids and lipopeptides, and facilitate species differentiation. MS hardware and techniques, for example, tandem MS, including diverse ion source/mass analyzer combinations are discussed. Relevant examples for microbial mixture characterization utilizing these combinations are provided. Chemometrics and bioinformatics methods and algorithms, including those applied to large scale MS data acquisition in microbial metaproteomics and MS imaging of biofilms, are highlighted. Select MS applications for polymicrobial culture analysis in environmental and clinical microbiology are reviewed as well.

中文翻译：

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通过质谱表征微生物混合物

在过去的10年中，MS在微生物学中的应用显着增加，部分原因是监管机构批准的用于快速识别临床感染的商用MALDI MS平台的激增。同时，随着质谱技术在“组学”领域的扩展，出现了基于质谱的新颖研究，以表征生物体和环境微生物组。对其他生物的复杂混合物中发现的微生物进行成功的表征仍然是研究人员和临床医生面临的主要挑战。在这里，我们回顾了MS在应对这一挑战方面的最新进展。这些方法包括样品制备方法和方案，并已建立，例如MALDI，以及较新的例如大气压力电离（API）技术。完整细胞的MALDI质谱图主要包含有关高度表达的用作生物标记物的管家蛋白的信息。API方法适用于小生物分子分析，例如磷脂和脂肽，并有助于物种分化。讨论了MS硬件和技术，例如串联MS，包括各种离子源/质量分析器组合。提供了利用这些组合表征微生物混合物的相关实例。重点介绍了化学计量学和生物信息学方法和算法，包括那些应用于微生物元蛋白质组学和生物膜MS成像的大规模MS数据采集的方法和算法。本文还对在环境和临床微生物学中用于微生物培养分析的部分MS应用进行了综述。