Brauge Thomas, Midelet-Bourdin Graziella, Soumet Christophe
Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health and Safety, Boulogne sur Mer, France.
RMT Chlean, Joint Technological Network: Hygienic Design of Production Lines and Equipment, Fougères, France.
Methods Mol Biol. 2019;1918:117-128. doi: 10.1007/978-1-4939-9000-9_9.
Foodborne pathogens are responsible of foodborne diseases and food poisoning and thus pose a great threat to food safety. These microorganisms can adhere to surface and form a biofilm composed of an extracellular matrix. This extracellular matrix protects bacterial cells from industrial environmental stress factors such as cleaning and disinfection operations. Moreover, during these environmental stresses, many bacterial species can enter a viable but nonculturable (VBNC) state. VBNC cells are characterized by a loss of cultivability on conventional bacteriological agar. This leads to an underestimation of total viable cells in environmental samples, and thus poses a risk for public health. In this chapter, we present a method to detect viable population of foodborne pathogens in industrial environmental samples using a molecular method with a combination of propidium monoazide (PMA) and quantitative PCR (qPCR) and a fluorescence microscopic method associated with the LIVE/DEAD BacLight™ viability stain.
食源性病原体是食源性疾病和食物中毒的罪魁祸首，因此对食品安全构成巨大威胁。这些微生物可以附着在表面并形成由细胞外基质组成的生物膜。这种细胞外基质可保护细菌细胞免受工业环境压力因素的影响，如清洁和消毒操作。此外，在这些环境压力下，许多细菌种类可以进入活的但不可培养（VBNC）状态。VBNC细胞的特征是在传统细菌学琼脂上丧失可培养性。这导致对环境样品中总活细胞的低估，从而对公众健康构成风险。在本章中，我们介绍了一种使用分子方法结合单叠氮丙锭（PMA）和定量PCR（qPCR）以及与LIVE/DEAD BacLight™活力染色相关的荧光显微镜方法来检测工业环境样品中食源性病原体活菌群的方法。
