Laboratory of Food Hygienic Chemistry, Division of Food Science and Biotechnology, Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University
九州大学大学院生物資源環境科学府生命機能科学専攻食糧科学工学コース食品衛生科学分野
Laboratory of Food Hygienic Chemistry, Division of Food Science and Biotechnology, Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University
九州大学大学院生物資源環境科学府生命機能科学専攻食糧科学工学コース食品衛生科学分野
Laboratory of Food Hygienic Chemistry, Division of Food Biotechnology, Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University
九州大学大学院生物資源環境科学府食品バイオ工学講座食品衛生化学分野
Laboratory of Food Hygienic Chemistry, Division of Food Biotechnology, Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University
九州大学大学院生物資源環境科学府食品バイオ工学講座食品衛生化学分野
Laboratory of Food Hygienic Chemistry, Division of Food Science and Biotechnology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University
九州大学大学院農学研究院生命機能科学専攻食糧科学工学コース食品衛生科学分野
Laboratory of Food Hygienic Chemistry, Division of Food Science and Biotechnology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University
九州大学大学院農学研究院生命機能科学専攻食糧科学工学コース食品衛生科学分野
Escherichia coli (E. coli) O157:H7 was detected by using a surface plasmon resonance (SPR) biosensor and two antibodies with different characters. The lower limit of detection of E. coli O157:H7 samples after pretreatments was determined by SPR. Seven pretreatment methods for preparing E. coli O157:H7 samples for SPR detection; beads disruption, sonication, and heat shock, osmotic shock, lysozyme, alkali, and boiling treatments were compared for SPR signal with untreated cells as a control. In the case of the antibody raised against intracellular substance, β–D–galactosidase (β–gal), was used for the detection, the lower limit of detection was 4.9×10^5 CFU/ml for both sonicated and alkali treated samples. The lower limit of detection was 8.3×10^6 CFU/ml for beads disrupted samples, and 8.2×10^8 CFU/ml for both lysozyme treated and untreated samples. In contrast, significant SPR signal was not obtained for heat shocked, osmotic shocked and boiled samples even at 8.2×10^8 CFU/ml. Sonication pretreatment improved the lower limit of detection for E. coli O157:H7 by three orders of magnitude compared with that of untreated sample when anti-β-gal antibody was used for detection by SPR biosensor. In the case of antibody raised against lipopolysaccharide (LPS), the cell surface substance, was used, sonicated E. coli O157:H7 sample was detected by SPR at 1.3×10^5 CFU/ml. The lower limit of detection was 1.1×10^6 CFU/ml for heat shocked, lysozyme treated, alkali treated, boiled and untreated samples, and 7.7×10^7 CFU/ml for beads disrupted samples, respectively. After the osmotic shock treatment, E. coli O157:H7 was not detected by SPR even at 2.1×10^8 CFU/ml. These results show that sonication was the most effective pretreatment method for the detection of E. coli O157:H7 by SPR using both antibodies recognizing intracellular β–gal, and cell surface LPS.
Mendeley出力
