title: Development of a simple and rapid diagnosis method for swine edema disease to specifically detect Stx2e protein by immunochromatographic test.
abstract: Edema disease in piglets is caused by Shiga toxin 2e (Stx2e)-producing Escherichia coli. However, there is currently no available Stx2e-specific immunochromatographic test strip to differentiate Stx2e from other types of Shiga toxin 2. In the present study, to develop an Stx2e-specific immunochromatographic test strip, we isolated nine different monoclonal antibody-producing hybridoma clones from Stx2e toxoid-immunized mice and confirmed that six antibodies were A subunit-specific whereas three antibodies were B subunit-specific. Only one A subunit-specific monoclonal antibody (45B2) was cross-reactive with prototype Stx2 (Stx2a) at the same sensitivity, but the remaining eight monoclonal antibodies were not. In immunochromatographic tests using the highly sensitive antibodies, test strips using some combinations of gold colloid-conjugated monoclonal antibody with the B subunit-specific monoclonal antibody on the membrane detected Stx2e, but not other types of Shiga toxin 2. These test strips had the ability to detect Stx2e in the culture supernatant of clinically isolated Stx2e gene-positive strains, but not in those of Stx2e gene-negative strains. These results indicate that our test strip is practical for the specific detection of Stx2e to diagnose swine edema disease.
authors.affiliation: Department of Microbiology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Arimitsu
authors.affiliation: Department of Microbiology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Department of Microbiology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Tsuji
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000992:Antitoxins / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D004488:Edema Disease of Swine / Q000175:diagnosis*; D051379:Mice; D012680:Sensitivity and Specificity; D022641:Shiga Toxin 2 / Q000032:analysis* / Q000276:immunology; D013552:Swine; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D000992:Antitoxins; D022641:Shiga Toxin 2
keywords: Shiga toxin; edema disease; immunochromatographic test; monoclonal antibody