RNA sequencing of foodborne pathogens

The study goal is to use direct metatranscriptome RNA-seq on Nanopore MinION to achieve real-time multiplex identification of viable pathogens in food. Specifically, this study reports an optimized universal Nanopore sample extraction and library preparation protocol applicable to both Gram-positive and Gram-negative pathogenic bacteria, demonstrated using a cocktail culture of E. coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes, which were selected based on their impact on economic loss or prevalence in recent outbreaks.

本研究旨在依托纳米孔MinION测序仪（Nanopore MinION）开展直接宏转录组RNA测序（metatranscriptome RNA-seq），以实现食品中活病原菌的实时多重鉴定。具体而言，本研究报道了一套适用于革兰氏阳性（Gram-positive）与革兰氏阴性（Gram-negative）病原菌的优化型通用纳米孔样本提取及文库制备流程，并通过大肠杆菌O157:H7、肠炎沙门氏菌及单核细胞增生李斯特菌的混合菌培养物验证了该流程的有效性；上述菌株均因其可造成重大经济损失或在近期暴发事件中高发而被选中。