title: Development of a rapid and simple immunochromatographic assay to identify Vibrio parahaemolyticus.
abstract: To rapidly and simply determine whether or not bacterial colonies growing on agar were Vibrio parahaemolyticus, we developed an immunochromatographic assay (VP-ICA) using two different monoclonal antibodies (designated mAb-VP34 and mAb-VP109) against the delta subunit of V. parahaemolyticus-F0F1 ATP synthase. The epitopes recognized by mAb-VP34 and mAb-VP109 were mapped to sequences of eight ((47)LLTSSFSA(54)) and six amino acid residues ((16)FDFAVD(21)), respectively. An amino acid sequence similarity search of the NCBI database using BLASTP showed that both epitopic amino acid sequences were present together only in V. parahaemolyticus. When 124 V. parahaemolyticus strains and 94 strains of 27 other Vibrio species or 35 non-Vibrio species were tested using the VP-ICA, the VP-ICA identified V. parahaemolyticus with 100% accuracy. The VP-ICA rapidly and simply identified the pathogen directly from a single agar colony within 30 min, indicating that VP-ICA will greatly reduce labor and time required to identify V. parahaemolyticus compared with conventional biochemical tests.
authors.affiliation: Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan. Electronic address: sakata@iph.pref.osaka.jp.
authors.forename: Junko
authors.initials: J
authors.lastname: Sakata
authors.affiliation: Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kawatsu
authors.affiliation: Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Iwasaki
authors.affiliation: Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kumeda
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D018604:Epitope Mapping / Q000379:methods*; D021122:Protein Subunits / Q000737:chemistry / Q000276:immunology; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000276:immunology; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D014736:Vibrio parahaemolyticus / Q000145:classification* / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D021122:Protein Subunits; D006180:Proton-Translocating ATPases
keywords: Epitope mapping; Identification; Immunochromatographic assay; Vibrio parahaemolyticus