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

被引:18
作者
Hayashi, Masahiro [1 ,2 ]
Natori, Tatsuya [2 ]
Kubota-Hayashi, Sayoko [2 ]
Miyata, Machiko [2 ]
Ohkusu, Kiyofumi [2 ]
Kawamoto, Keiko [3 ]
Kurazono, Hisao [3 ]
Makino, Souichi [4 ]
Ezaki, Takayuki [2 ]
机构
[1] Gifu Univ, Life Sci Res Ctr, Div Anaerobe Res, Gifu 5011194, Japan
[2] Gifu Univ, Dept Microbiol, Grad Sch Med, Gifu 5011194, Japan
[3] Obihiro Univ Agr & Vet Med Inada Cho, Dept Anim & Food Hyg, Div Food Hyg, Obihiro, Hokkaido 0808555, Japan
[4] Kyoto Seibo Coll, Dept Domest Sci, Kyoto 6120878, Japan
关键词
CAMPYLOBACTER-JEJUNI; ESCHERICHIA-COLI; SALMONELLA SPP; LISTERIA-MONOCYTOGENES; PUBLIC-HEALTH; IDENTIFICATION; ASSAY; QUANTIFICATION; AMPLIFICATION; FECES;
D O I
10.1155/2013/295050
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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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页数:10
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