ADRIA Développement-UMT 08.3PHYSI'Opt, Creac'h Gwen, 29196 Quimper, France.
Food Microbiol. 2011 Aug;28(5):848-61. doi: 10.1016/j.fm.2011.02.008. Epub 2011 Apr 1.
Molecular methods are being increasingly applied to detect, quantify and study microbial populations in food or during food processes. Among these methods, PCR-based techniques have been the subject of considerable focus and ISO guidelines have been established for the detection of food-borne pathogens. More particularly, real-time quantitative PCR (qPCR) is considered as a method of choice for the detection and quantification of microorganisms. One of its major advantages is to be faster than conventional culture-based methods. It is also highly sensitive, specific and enables simultaneous detection of different microorganisms. Application of reverse-transcription-qPCR (RT-qPCR) to study population dynamics and activities through quantification of gene expression in food, by contrast with the use of qPCR, is just beginning. Provided that appropriate controls are included in the analyses, qPCR and RT-qPCR appear to be highly accurate and reliable for quantification of genes and gene expression. This review addresses some important technical aspects to be considered when using these techniques. Recent applications of qPCR and RT-qPCR in food microbiology are given. Some interesting applications such as risk analysis or studying the influence of industrial processes on gene expression and microbial activity are reported.
分子方法正越来越多地应用于检测、定量和研究食品中的微生物种群或在食品加工过程中的微生物种群。在这些方法中，基于 PCR 的技术一直是相当关注的焦点，并且已经为食源性致病菌的检测制定了 ISO 指南。更具体地说，实时定量 PCR（qPCR）被认为是检测和定量微生物的首选方法。其主要优点之一是比传统基于培养的方法更快。它还具有高度的敏感性、特异性，并能够同时检测不同的微生物。通过在食品中定量表达基因来研究种群动态和活性的逆转录 qPCR（RT-qPCR）的应用，与 qPCR 相比，刚刚开始。只要在分析中包括适当的对照，qPCR 和 RT-qPCR 似乎非常准确和可靠，可用于定量基因和基因表达。本文讨论了使用这些技术时需要考虑的一些重要技术方面。给出了 qPCR 和 RT-qPCR 在食品微生物学中的最新应用。报告了一些有趣的应用，如风险分析或研究工业过程对基因表达和微生物活性的影响。
