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食品环境中食源性致病菌的 PMA-qPCR 和显微镜观察法的活力检测。

Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopy Observation.

机构信息

Laboratory for food safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Boulogne sur Mer, France.

French Agency for Food, Environmental and Occupational Health & Safety, Laboratory for food safety, Boulogne sur Mer, France.

出版信息

Methods Mol Biol. 2025;2852:33-46. doi: 10.1007/978-1-0716-4100-2_3.

DOI:10.1007/978-1-0716-4100-2_3
PMID:39235735
Abstract

Foodborne pathogens are responsible for 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 matrix protects bacterial cells from industrial environmental stress factors such as cleaning and disinfection operations. Moreover, during these environmental stresses, many bacterial species can be entered in a viable but nonculturable (VBNC) state. VBNC cells are characterized by an active metabolism and a loss of cultivability on conventional bacteriological agar. This leads to an underestimation of total viable cells in environmental samples and thus may pose 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 combining propidium monoazide (PMA) and quantitative PCR (qPCR) and a fluorescence microscopic method associated with the LIVE/DEAD BacLight™ viability stain.

摘要

食源性致病菌可导致食源性疾病和食物中毒,对食品安全构成极大威胁。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。该基质可保护细菌细胞免受工业环境胁迫因素(如清洗和消毒操作)的影响。此外,在这些环境胁迫下,许多细菌物种可以进入存活但非可培养(VBNC)状态。VBNC 细胞的特征是新陈代谢活跃,在常规细菌琼脂上丧失可培养性。这导致环境样本中总活菌的低估,从而可能对公共健康构成威胁。在本章中,我们提出了一种使用结合了吖啶橙(PMA)和定量 PCR(qPCR)的分子方法以及与 LIVE/DEAD BacLight™活菌染色剂相关的荧光显微镜方法来检测工业环境样本中食源性致病菌存活种群的方法。

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本文引用的文献

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Understanding the reaction balances behind the viability PCR protocols based on photoreactive dyes.理解基于光反应性染料的活力 PCR 方案背后的反应平衡。
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Enumeration of viable Listeria monocytogenes cells by real-time PCR with propidium monoazide and ethidium monoazide in the presence of dead cells.在存在死细胞的情况下,使用单叠氮化丙锭和单叠氮化乙锭通过实时聚合酶链反应对单核细胞增生李斯特氏菌活细胞进行计数。
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