A new protocol to detect multiple foodborne pathogens with PCR dipstick DNA chromatography after a six-hour enrichment culture in a broad-range food pathogen enrichment broth

一种利用PCR试纸DNA层析法检测多种食源性病原体的新方案,该方法是在广谱食源性病原体增菌培养液中进行6小时增菌培养后进行的。

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作者:Masahiro Hayashi,Tatsuya Natori,Sayoko Kubota-Hayashi,Machiko Miyata,Kiyofumi Ohkusu,Keiko Kawamoto,Hisao Kurazono,Souichi Makino,Takayuki Ezaki

Abstract

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.

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