title: [Molecular Detection Methods for Vibrio parahaemolyticus in Seafood].
abstract: To detect Vibrio parahaemolyticus in seafood, we evaluated efficient combinations of molecular methods with DNA extraction methods using heat extraction and alkaline heat extraction, and PCR, real-time PCR and loop-mediated isothermal amplification (LAMP) assays were performed targeting V parahaemolyticus species-specific genes (tlh and rpoD) and pathogenic factors genes (tdh and trh). The species-specific genes were detected in all combinations of two strains (a tdh * trh1-positive strain and a trh2-positive strain), two kinds of shellfish (oyster and bloody clams) and molecular methods with tlh-real time PCR or rpoD-LAMP assays with DNA of alkaline heat extraction at 85-145cfu/test level. tdh was detected in both seafoods with real time PCR assay with DNA of heat extraction at 85cfu/test level, and detected with the LAMP and real time PCR assays with DNA of alkaline heat extraction at 85cfu/test level. Detection of both trh1 and trh2 with the PCR assay with DNA of alkaline heat extraction was comparatively high though trh2 was detected with the LAMP assay with DNA of alkaline heat extraction at 145cfu/test level. It, however, is necessary to investigate more sensitive trh-detection methods. In this study, the results indicated that tlh-real time PCR or rpoD-LAMP, tdh-real time PCR and tdh-LAMP assays with DNA of alkaline heat extraction are relatively-sensitive methods to detect V. parahaemolyticus in seafood.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Nishio
authors.forename: Kayoko
authors.initials: K
authors.lastname: Ohtsuka
authors.forename: Midori
authors.initials: M
authors.lastname: Oda
authors.forename: Kanji
authors.initials: K
authors.lastname: Sugiyama
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Hara-Kudo
mesh_terms: D000818:Animals; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D017747:Seafood / Q000382:microbiology*; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification*