title: Development of a Surface Plasmon Resonance-Based Immunosensor for Detection of 10 Major O-Antigens on Shiga Toxin-Producing Escherichia coli, with a Gel Displacement Technique To Remove Bound Bacteria.
abstract: A surface plasmon resonance-based immunosensor (SPR-immunosensor) was developed for the detection of Shiga toxin-producing Escherichia coli (STEC) belonging to the O-antigen groups O26, O91, O103, O111, O115, O121, O128, O145, O157, and O159. The polyclonal antibodies (PoAbs) generated against each of the STEC O-antigen types in rabbits were purified and were immobilized on the sensor chip at 0.5 mg/mL. The limit of detection for STEC O157 by the SPR-immunosensor was found to be 6.3 × 10(4) cells for 75 s. Each of the examined 10 O-antigens on the STECs was detected by the corresponding PoAb with almost no reaction to the other PoAbs. The detected STECs were sufficiently removed from the PoAbs using gelatin or agarose gel without deactivation of the PoAbs, enabling repeatable use of the sensor chip. The developed SPR-immunosensor can be applied for the detection of multiple STEC O-antigens. Furthermore, the new antigen removal technique using the gel displacement approach can be utilized with various immunosensors to improve the detection of pathogens in clinical and public health settings.
authors.affiliation: Advanced Science, Technology & Management Research Institute of Kyoto , Shimogyo-ku, Kyoto 600-8813, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Yamasaki
authors.affiliation: Advanced Science, Technology & Management Research Institute of Kyoto , Shimogyo-ku, Kyoto 600-8813, Japan.|Research & Development Division, HORIBA, Ltd., Minami-ku, Kyoto 601-8510, Japan.
authors.forename: Shiro
authors.initials: S
authors.lastname: Miyake
authors.affiliation: Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine , Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nakano
authors.affiliation: Research & Development Division, HORIBA, Ltd., Minami-ku, Kyoto 601-8510, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Morimura
authors.affiliation: Advanced Science, Technology & Management Research Institute of Kyoto , Shimogyo-ku, Kyoto 600-8813, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hirakawa
authors.affiliation: Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine , Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Nagao
authors.affiliation: Department of Microbiology, Kobe Institute of Health , Chuo-ku, Kobe 650-0046, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Iijima
authors.affiliation: Department of Food and Nutrition, Kyoto Women's University , Higashiyama-ku, Kyoto 605-8501, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Narita
authors.affiliation: Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine , Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ichiyama
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D019453:Escherichia coli O157 / Q000378:metabolism*; D007118:Immunoassay; D057230:Limit of Detection; D046228:Microarray Analysis; D019081:O Antigens / Q000032:analysis* / Q000276:immunology; D022621:Shiga Toxin / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D055910:Antibodies, Immobilized; D019081:O Antigens; D022621:Shiga Toxin