title: Development of an indirect competitive ELISA for the detection of ABO blood group antigens.
abstract: We developed an indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection of ABO blood group antigens in human samples; in particular for blood stains. ABO blood group antigens conjugated to polyacrylamide were used for immobilized antigen. ABO blood group antigens were extracted from blood stains using a novel method involving pre-incubation with proteinase K (PK), followed by heat treatment. The extracts (analytes) were combined with either anti-A or -B monoclonal antibodies (mAbs), and added directly to the antigen-coated wells. The anti-A and -B mAbs were captured by either ABO blood group antigens present in the analyte or by the immobilized blood group antigens. Peroxidase-conjugated anti-mouse IgM antibody was used to detect anti-A and -B mAbs complexed with immobilized blood group antigens, and a colorimetric reaction using o-phenylenediamine/H2O2 used for its measurement. The ELISA developed in this study was able to detect blood group antigens in blood, saliva and blood stains. The detection limit for unknown blood, saliva and blood stain were determined as 1:200, 1:32 and 1:16. Overall the ABO blood grouping ELISA can be used with relative ease for the high throughput screening of biological samples for the detection of ABO blood group antigens.
authors.affiliation: Science of Biological Resources, United Graduate School of Agricultural Sciences, Gifu University, Gifu 501-1193, Japan; Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan. Electronic address: naokiutd_1977@ybb.ne.jp.
authors.forename: Naoki
authors.initials: N
authors.lastname: Takada
authors.affiliation: Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan.
authors.forename: Chikahiro
authors.initials: C
authors.lastname: Mori
authors.affiliation: Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan.
authors.forename: Mizuho
authors.initials: M
authors.lastname: Iida
authors.affiliation: Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Takai
authors.affiliation: Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Takayama
authors.affiliation: Forensic Science Laboratory, Gifu Prefectural Police Headquarters, Gifu 500-8501, Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Faculty of Applied Biological Sciences, Gifu University, Gifu 501-1193, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: Faculty of Applied Biological Sciences, Gifu University, Gifu 501-1193, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Takamizawa
mesh_terms: D000017:ABO Blood-Group System / Q000097:blood*; D000911:Antibodies, Monoclonal / Q000097:blood; D000918:Antibody Specificity; D001788:Blood Grouping and Crossmatching / Q000379:methods; D001801:Blood Stains; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D049429:Forensic Pathology; D006801:Humans; D012463:Saliva / Q000276:immunology
chemical_list: D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal
keywords: ABO blood grouping; Blood group antigen; Indirect competitive ELISA; Monoclonal antibody