Examination of quick inspection method of diarrhea primitive Escherichia coli in meat and feces by polymerase chain reaction.
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概要
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A direct polymerase chain reaction method to EC cultured broth (named EC-PCR) for screening detection of enteropathogenic <I>E. coli</I> (ETEC, VTEC and EIEC) in foods and feces is described.<BR>Eleven <I>E. coli</I> strains belonging to three groups (4ETEC: 06, 025, 0128, 0148; 4VTEC: 026, 0111, 0145, 0157; and 3EIEC: 028ac, 0124, 0164) were used for this study. Four pairs of oligonucleotide primers homologous to LT, ST, VT and EIEC genes were used in combination. The culture condition at 37-43°C for 16-20 h in EC broth was most suitable for the EC-PCR method, and no cross readings were observed with other bacteria, substances in meats or feces.<BR>After a 20-h enrichment step, it was possible to detect fewer than 10 to 10<SUP>2</SUP> bacteria per g of the aritificially inoculated meat. <I>E. coli</I> was easily detected because of low contamination with other bacteria which disturb the detection of <I>E. coli</I>. However, in feces, it ranged from 10<SUP>3</SUP> to 10<SUP>4</SUP> cfu per g. The large number of bacteria with feces are the main limiting factor of the EC-PCR detection assay. All <I>E. coli</I> strains examined were detected from all the enrichment cultures on both the EC-PCR and the culture methods. In two sporadic cases and two food poisoning cases, the enteropathogenicity of <I>E. coli</I> isolates from patients was rapidly judged by the EC-PCR method.<BR>These findings were consistent with those of the culture method. Thus, the findings suggest that the EC-PCR method is a suitable, sensitive and rapid method for detection of the potentially enteropathogenic <I>E. coli</I>.
- 日本食品微生物学会の論文
日本食品微生物学会 | 論文
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