食中毒細菌5種の一斉増菌同時検出

概要

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A single broth enrichment based polymerase chain reaction (PCR) assay for simultaneous detection of five major food poisoning bacteria, Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, and Bacillus cereus (emetic) has been developed. All of target bacteria were simultaneously inoculated in a single broth, tryptic soy broth (TSB) supplement with 1.5% NaCl. After 18-h incubation at 37℃, viable cell counts of all bacteria increased to enough amount of cell density (more than 10^4 CFU/ml) for DNA extraction used subsequent PCR assay. PCR was performed with specific primer sets for each bacterium. The targeted genes used were: ipaB of salmonella spp, eaeA, stxl, and stx2 of E. coli O157:H7, tl, tdh, and trh of V. parahaemolyticus, nuc of S. aureus, and crs of B. cereus. The developed method could detect all target bacteria at initial inoculation level of 1 to 10 CFU/25g of food samples (chickin, milk, and read-to-eat vegetable) within 24 h without any inhibition from food ingredients or bacteria derived from food.
九州大学の論文