Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 19 Chlorine Gardens, Belfast, BT9 5DL, Northern Ireland, UK.
Appl Microbiol Biotechnol. 2020 May;104(10):4281-4288. doi: 10.1007/s00253-020-10542-x. Epub 2020 Mar 26.
The ability to rapidly detect viable pathogens in food is important for public health and food safety reasons. Culture-based detection methods, the traditional means of demonstrating microbial viability, tend to be laborious, time consuming and slow to provide results. Several culture-independent methods to detect viable pathogens have been reported in recent years, including both nucleic acid-based (PCR combined with use of cell viability dyes or reverse-transcriptase PCR to detect messenger RNA) and phage-based (plaque assay or phage amplification and lysis plus PCR/qPCR, immunoassay or enzymatic assay to detect host DNA, progeny phages or intracellular components) methods. Some of these newer methods, particularly phage-based methods, show promise in terms of speed, sensitivity of detection and cost compared with culture for food testing. This review provides an overview of these new approaches and their food testing applications, and discusses their current limitations and future prospects in relation to detection of viable pathogens in food. KEY POINTS: • Cultural methods may be 'gold standard' for assessing viability of pathogens, but they are too slow. • Nucleic acid-based methods offer speed of detection but not consistently proof of cell viability. • Phage-based methods appear to offer best alternative to culture for detecting viable pathogens.
快速检测食品中存活病原体的能力对于公共卫生和食品安全至关重要。基于培养的检测方法是证明微生物存活能力的传统手段,但往往繁琐、耗时且结果缓慢。近年来,已经报道了几种不依赖培养的方法来检测存活病原体,包括基于核酸的方法(PCR 结合使用细胞活力染料或逆转录 PCR 检测信使 RNA)和基于噬菌体的方法(噬菌斑测定或噬菌体扩增和裂解加 PCR/qPCR、免疫测定或酶测定检测宿主 DNA、后代噬菌体或细胞内成分)。与食品检测中的培养相比,这些较新方法中的一些方法,特别是基于噬菌体的方法,在速度、检测灵敏度和成本方面具有很大的优势。本文综述了这些新方法及其在食品检测中的应用,并讨论了它们在检测食品中存活病原体方面的当前局限性和未来前景。要点:
培养方法可能是评估病原体存活能力的“金标准”,但速度太慢。
基于核酸的方法提供了检测速度,但不能始终证明细胞存活。
基于噬菌体的方法似乎是检测存活病原体的培养法的最佳替代方法。