Centre for Advanced Sensory Science, School of Exercise and Nutrition Sciences, Deakin University, Melbourne, VIC, Australia.
Department of Chemistry and Biotechnology, Swinburne University of Technology, Melbourne, VIC, Australia.
Methods Mol Biol. 2021;2220:17-29. doi: 10.1007/978-1-0716-0982-8_2.
Listeria monocytogenes is a major food-borne pathogen and causative agent of a fatal disease, listeriosis. Stringent regulatory guidelines and zero tolerance policy toward this bacterium necessitate rapid, accurate, and reliable methods of identification and subtyping. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) has recently become a method of choice for routine identification of pathogens in clinical settings and has largely replaced biochemical assays. Identification relies on well-curated databases such as SARAMIS. Extensive use of SARAMIS to generate consensus mass spectra, in conjunction with statistical analysis, such as partial least square-discriminant analysis and hierarchical cluster analysis, is useful in subtyping bacteria. While MALDI-ToF MS has been extensively used for pathogen detection, its application in bacterial subtyping has been limited. The protocol describes a MALDI-ToF MS workflow as a single tool for simultaneous identification and subtyping of L. monocytogenes directly from solid culture medium.
单增李斯特菌是一种主要的食源性致病菌，也是李斯特菌病的病原体。为了应对这一致病菌，监管法规非常严格，实行零容忍政策，这就需要快速、准确、可靠的鉴定和分型方法。基质辅助激光解吸电离飞行时间质谱（MALDI-ToF MS）最近已成为临床环境中常规鉴定病原体的首选方法，在很大程度上取代了生化检测方法。鉴定依赖于精心维护的数据库，如 SARAMIS。广泛使用 SARAMIS 来生成共识质谱，结合统计分析，如偏最小二乘判别分析和层次聚类分析，有助于对细菌进行分型。虽然 MALDI-ToF MS 已广泛用于病原体检测，但在细菌分型方面的应用有限。该方案描述了 MALDI-ToF MS 工作流程，可直接从固体培养介质中作为一种单一工具，同时对单增李斯特菌进行鉴定和分型。
