Assaf A, Cordella C B Y, Thouand G
University of Nantes, UMR CNRS GEPEA 6144, CBAC, 18 Boulevard Gaston Defferre, CS50020, 85035, Nantes, La Roche sur Yon, France.
Anal Bioanal Chem. 2014 Aug;406(20):4899-910. doi: 10.1007/s00216-014-7909-2. Epub 2014 Jun 8.
Food safety is a major concern for suppliers in the food chain to ensure the safety of their products. The identification procedure requested by norms is tedious, and it often requires systematic controls and qualified staff to perform the necessary analyses. Raman spectroscopy offers new opportunities to rapidly and efficiently ascertain the presence of pathogens in samples. Nevertheless, this technique requires a standardized procedure to be applied in the industrial context. Our study shows that the variability between spectral fingerprints is related to the physiological state of the microbial species and the growth phase of the bacteria plays a crucial role in its identification by Raman spectroscopy. To improve the discrimination between closely related bacterial species, a procedure based on the selection of bacterial spectra in the exponential growth phase was proposed. Different ways to introduce Raman spectroscopy in the ISO 6579:2002 standards are also proposed from the entire process to a shorter protocol. In the latter case, the identification of bacterial colonies after the selective enrichment step was proposed with the advantages of this path in terms of simplicity and rapidity (analysis time is reduced up to 50 h from the 100 h required by the standard). The protocol validated using six food categories from industrial partners have presented a good correlation by confirmation with other laboratory classical methods. In the future, this procedure could be introduced to the control system of the food production chain with a reliable database for various microorganisms encountered in this field.
食品安全是食品链供应商确保其产品安全的主要关注点。规范要求的鉴定程序繁琐,通常需要系统的控制和合格的人员来进行必要的分析。拉曼光谱法为快速有效地确定样品中病原体的存在提供了新机会。然而,该技术需要在工业环境中应用标准化程序。我们的研究表明,光谱指纹之间的变异性与微生物物种的生理状态有关,并且细菌的生长阶段在通过拉曼光谱法进行鉴定中起着关键作用。为了提高密切相关细菌物种之间的区分度,提出了一种基于选择指数生长期细菌光谱的程序。还从整个过程到更短的方案提出了将拉曼光谱法引入ISO 6579:2002标准的不同方法。在后一种情况下,提出了在选择性富集步骤后鉴定细菌菌落的方法,该方法在简单性和快速性方面具有优势(分析时间从标准要求的100小时减少到50小时)。使用来自工业合作伙伴的六种食品类别验证的方案通过与其他实验室经典方法的确认呈现出良好的相关性。未来,该程序可以引入食品生产链的控制系统,并建立一个可靠的数据库,用于该领域遇到的各种微生物。