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

نویسندگان

  • Masahiro Hayashi
  • Tatsuya Natori
  • Sayoko Kubota-Hayashi
  • Machiko Miyata
  • Kiyofumi Ohkusu
  • Keiko Kawamoto
  • Hisao Kurazono
  • Souichi Makino
  • Takayuki Ezaki
چکیده

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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عنوان ژورنال:

دوره 2013  شماره 

صفحات  -

تاریخ انتشار 2013