Center for Advanced Measurement Science, National Institute of Metrology, Beijing, China.
Research Center for Ginseng Genetic Resources Development and Utilization, Jilin Agricultural University, Changchun, China.
Foodborne Pathog Dis. 2020 Jul;17(7):447-458. doi: 10.1089/fpd.2019.2748. Epub 2020 Jan 31.
, a highly virulent food-borne pathogen transmitted through food, can cause severe infectious diseases in a large number of people through a single outbreak, due to its low infective doses. In this study, a flow cytometry (FCM)-based method was developed for the rapid detection of single viable cells with dual staining of fluorescein isothiocyanate (FITC)-labeled anti- antibody and propidium iodide (PI) dyes. The FCM-based method includes 6 h of pre-enrichment, 40 min of target cell isolation, and 20 min of dual staining and FCM analysis. The developed method demonstrated high specificity for the detection of 23 strains and 22 food-borne pathogenic non- strains. Furthermore, the analyses of 30 samples of milk powder artificially contaminated with single cells, 123 samples of retail milk powder, and 6 samples of -positive milk powder were performed by the FCM-based as well as traditional plate-based methods for testing the efficiency of the methods. The two methods yielded similar results for the detection of pathogens in all milk powder samples. In conclusion, the developed FCM-based method was found to be efficient in detecting single viable cells in milk powder within 7 h. The proposed dual-color FITC assay combined with pre-enrichment offers a great potential for the rapid and sensitive detection of other pathogens in dairy products.
, 一种通过食物传播的高毒力食源性病原体，由于其感染剂量低，通过单次暴发即可在大量人群中引起严重的传染病。在本研究中，建立了一种基于流式细胞术（FCM）的方法，使用荧光素异硫氰酸酯（FITC）标记的抗-抗体和碘化丙啶（PI）染料双重染色，用于快速检测单个活的 细胞。该基于 FCM 的方法包括 6 小时的预富集、40 分钟的目标细胞分离以及 20 分钟的双重染色和 FCM 分析。所建立的方法对 23 株 和 22 株食源性病原体非- 菌株的检测表现出高度特异性。此外，通过基于 FCM 的方法和传统的平板方法对人工污染有单个 细胞的 30 份奶粉样品、123 份零售奶粉样品和 6 份阳性奶粉样品进行了分析，以测试方法的效率。两种方法在所有奶粉样品中对病原体的检测均产生了相似的结果。总之，所建立的基于 FCM 的方法在 7 小时内能够有效检测奶粉中的单个活的 细胞。所提出的双重颜色 FITC 测定法与预富集相结合，为乳制品中其他病原体的快速灵敏检测提供了巨大潜力。
