Hayashi Masahiro, Natori Tatsuya, Kubota-Hayashi Sayoko, Miyata Machiko, Ohkusu Kiyofumi, Kawamoto Keiko, Kurazono Hisao, Makino Souichi, Ezaki Takayuki
Division of Anaerobe Research, Life Science Research Center, Gifu University, 1-1 Yanagido, Gifu 501-1194, Japan ; Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan.
Biomed Res Int. 2013;2013:295050. doi: 10.1155/2013/295050. Epub 2013 Dec 2.
A quick foodborne pathogen screening method after six-hour enrichment culture with a broad-range food pathogen enrichment broth is described. Pathogenic factors of Salmonella enterica, Shigella spp., enteroinvasive Escherichia coli, and enterohemorrhagic E. coli are amplified with a cocktail primer and rapid polymerase chain reaction (PCR), which finishes amplification in 30 min. The PCR amplicon was differentiated with a dipstick DNA chromatography assay in 5-10 min. Starting from a four- to six-hour enrichment culture, this assay was finished within 45 min. Detection sensitivity of this protocol was less than 2.5 CFU/25 g for S. enterica and 3.3 CFU/25 g for enterohemorrhagic E. coli in spiked ground meat experiments.
本文描述了一种使用广谱食品病原体富集肉汤进行六小时富集培养后的快速食源性病原体筛查方法。用混合引物和快速聚合酶链反应(PCR)扩增肠炎沙门氏菌、志贺氏菌属、肠侵袭性大肠杆菌和肠出血性大肠杆菌的致病因子,该反应在30分钟内完成扩增。PCR扩增产物通过试纸条DNA色谱分析在5-10分钟内进行鉴别。从四至六小时的富集培养开始,该检测在45分钟内完成。在加标绞肉实验中,该方法对肠炎沙门氏菌的检测灵敏度低于2.5 CFU/25 g,对肠出血性大肠杆菌的检测灵敏度低于3.3 CFU/25 g。