Center for Biomolecular Nanotechnologies, Fondazione Istituto Italiano di Tecnologia @UniLe, via Barsanti 1, I-73010 Arnesano, Lecce, Italy.
Food Microbiol. 2013 Aug;35(1):10-4. doi: 10.1016/j.fm.2013.02.010. Epub 2013 Mar 1.
The development of fast, reliable and culture-independent molecular tools to detect bacteria producing biogenic amines deserves the attention of research and ultimately of the food industry in order to protect consumers' health. Here we present the application of a simple, low-cost, fast and sensitive method to perform microdroplet-based multiplex PCR, directly on a food matrix, for the simultaneous detection of bacterial genes involved in biogenic amine biosynthesis. After inoculating wine with Lactobacillus brevis IOEB 9809, cell lysis and DNA amplification are performed in one single step, without preliminary nucleic acid extraction or purification treatments. The assay is performed in about 30 min, requiring 150 nL of starting sample and it enables the detection of down to 15 bacterial cells. With respect to traditional culture techniques, the speed, the simplicity and the cheapness of this procedure allow an effective monitoring of microbial cells during food-making and processing.
为了保护消费者的健康，开发快速、可靠且与文化无关的分子工具来检测产生生物胺的细菌，值得研究人员和食品行业的关注。在这里，我们展示了一种简单、低成本、快速和敏感的方法在食品基质上直接进行基于微滴的多重 PCR，用于同时检测参与生物胺生物合成的细菌基因。在将葡萄酒接种到乳杆菌 IOEB 9809 后，在一个步骤中完成细胞裂解和 DNA 扩增，无需进行核酸提取或纯化处理。该测定在大约 30 分钟内完成，需要 150nL 的起始样品，能够检测到低至 15 个细菌细胞。与传统的培养技术相比，该方法的速度、简单性和廉价性允许在食品制作和加工过程中对微生物细胞进行有效监测。
