Jadhav Snehal R, Shah Rohan M, Karpe Avinash V, Morrison Paul D, Kouremenos Konstantinos, Beale David J, Palombo Enzo A
Department of Chemistry and Biotechnology, Swinburne University of Technology, Melbourne, VIC, Australia.
Land and Water, Commonwealth Scientific and Industrial Research Organisation, Brisbane, QLD, Australia.
Front Microbiol. 2018 Dec 18;9:3132. doi: 10.3389/fmicb.2018.03132. eCollection 2018.
Considering the short shelf-life of certain food products such as red meat, there is a need for rapid and cost-effective methods for pathogen detection. Routine pathogen testing in food laboratories mostly relies on conventional microbiological methods which involve the use of multiple selective culture media and long incubation periods, often taking up to 7 days for confirmed identifications. The current study investigated the application of omics-based approaches, proteomics using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-ToF MS) and metabolomics using gas chromatography-mass spectrometry (GC-MS), for detection of three red meat pathogens - , and O157:H7. Species-level identification was achieved within 18 h for and O157:H7 and 30 h for using MALDI-ToF MS analysis. For the metabolomics approach, metabolites were extracted directly from selective enrichment broth samples containing spiked meat samples (obviating the need for culturing on solid media) and data obtained using GC-MS were analyzed using chemometric methods. Putative biomarkers relating to , and O157:H7 were observed within 24, 18, and 12 h, respectively, of inoculating meat samples. Many of the identified metabolites were sugars, fatty acids, amino acids, nucleosides and organic acids. Secondary metabolites such as cadaverine, hydroxymelatonin and 3,4-dihydroxymadelic acid were also observed. The results obtained in this study will assist in the future development of rapid diagnostic tests for these important foodborne pathogens.
考虑到某些食品(如红肉)的保质期较短,因此需要快速且经济高效的病原体检测方法。食品实验室的常规病原体检测大多依赖于传统的微生物学方法,这些方法需要使用多种选择性培养基并经过较长的培养期,通常需要长达7天才能得到确诊结果。本研究调查了基于组学的方法的应用,即使用基质辅助激光解吸电离飞行时间质谱(MALDI-ToF MS)的蛋白质组学和使用气相色谱 - 质谱(GC-MS)的代谢组学,用于检测三种红肉病原体——[此处原文缺失病原体名称]、[此处原文缺失病原体名称]和O157:H7。使用MALDI-ToF MS分析,[此处原文缺失病原体名称]和O157:H7在18小时内实现了种水平鉴定,[此处原文缺失病原体名称]在30小时内实现了种水平鉴定。对于代谢组学方法,直接从含有加标肉样的选择性富集肉汤样品中提取代谢物(无需在固体培养基上培养),并使用化学计量学方法分析通过GC-MS获得的数据。在接种肉样后的24小时、18小时和12小时内,分别观察到了与[此处原文缺失病原体名称]、[此处原文缺失病原体名称]和O157:H7相关的推定生物标志物。许多鉴定出的代谢物为糖类、脂肪酸、氨基酸、核苷和有机酸。还观察到了尸胺、羟基褪黑素和3,4 - 二羟基马德酸等次生代谢物。本研究获得的结果将有助于未来针对这些重要食源性病原体的快速诊断测试的开发。