title: Production and characterization of a novel monoclonal antibody against Vibrio parahaemolyticus F0F1 ATP synthase's delta subunit and its application for rapid identification of the pathogen.
abstract: We raised monoclonal antibodies (MAbs) against Vibrio parahaemolyticus cell extracts. One of the MAbs, designated MAb-VP34, reacted in enzyme-linked immunosorbent assays (ELISAs) with 140 V. parahaemolyticus strains, regardless of serotype or origin. MAb-VP34 did not detectably react with 96 strains belonging to 27 other Vibrio species (except for Vibrio natriegens) or with 29 non-Vibrio species. These results show that MAb-VP34 is highly specific for V. parahaemolyticus. Western blotting and mass spectrometry analyses revealed that MAb-VP34 recognized V. parahaemolyticus F(0)F(1) ATP synthase's delta subunit. Using MAb-VP34, a rapid and simple immunodot blotting assay (VP-Dot) was developed to determine whether bacterial colonies growing on selective agar, represented V. parahaemolyticus. To evaluate VP-Dot, 20 V. parahaemolyticus strains and 19 non-related strains were tested. The results indicated that VP-Dot is a reliable tool for identification of V. parahaemolyticus colonies. The simple VP-Dot procedure took 40min, indicating that the MAb-VP34 based immunological method will greatly reduce labor, time, and costs required to verify V. parahaemolyticus colonies as compared with the conventional biochemical test.
authors: Division of Bacteriology, Osaka Prefectural Institute of Public Health, 1-3-69, Nakamichi, Higashinari-ku, Osaka 537-0025, Japan. sakata@iph.pref.osaka.jp
authors: Junko
authors: J
authors: Sakata
authors: Kentaro
authors: K
authors: Kawatsu
authors: Tadashi
authors: T
authors: Iwasaki
authors: Katsuhiro
authors: K
authors: Tanaka
authors: Shigeo
authors: S
authors: Takenaka
authors: Yuko
authors: Y
authors: Kumeda
authors: Hiroshi
authors: H
authors: Kodama
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001426:Bacterial Proteins / Q000276:immunology*; D015373:Bacterial Typing Techniques / Q000295:instrumentation / Q000379:methods*; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D021122:Protein Subunits / Q000276:immunology; D006180:Proton-Translocating ATPases / Q000276:immunology*; D017747:Seafood / Q000382:microbiology; D014735:Vibrio Infections / Q000382:microbiology*; D014736:Vibrio parahaemolyticus / Q000145:classification / Q000201:enzymology / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D021122:Protein Subunits; D006180:Proton-Translocating ATPases