title: Evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Vibrio parahaemolyticus in naturally contaminated seafood samples.
abstract: We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of seafood samples naturally contaminated with Vibrio parahaemolyticus. A total of 171 seafood samples enriched in alkaline peptone water (APW) were assessed by LAMP assay and conventional culture methods, which consist of a combination of APW enrichment culture and plating onto CHROMagar Vibrio and TCBS agars. Compared with V. parahaemolyticus isolation using the conventional culture test, LAMP results showed 100% (30/30) and 90.8% (128/141) sensitivity and specificity, respectively. The conventional culture test required more than 3 days to isolate and identify V. parahaemolyticus in the APW enrichment culture. In contrast, the LAMP assay was markedly faster, requiring less than 60 min from the beginning of DNA extraction to final detection of V. parahaemolyticus. In total, the LAMP assay required 17-19 h from the beginning of enrichment culture to final determination. This is the first report of the LAMP assay for rapid screening of seafood samples naturally contaminated by V. parahaemolyticus.
authors.affiliation: Laboratory of Veterinary Public Health, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuen Kibanadai-nishi, Miyazaki 889-2192, Japan. yamazakiw@cc.miyazaki-u.ac.jp
authors.forename: Wataru
authors.initials: W
authors.lastname: Yamazaki
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kumeda
authors.forename: Ryoko
authors.initials: R
authors.lastname: Uemura
authors.forename: Naoaki
authors.initials: N
authors.lastname: Misawa
mesh_terms: D000818:Animals; D005399:Fishes / Q000382:microbiology*; D005506:Food Contamination / Q000032:analysis; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D049897:Pectinidae / Q000382:microbiology*; D033561:Penaeidae / Q000382:microbiology*; D017747:Seafood / Q000382:microbiology*; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification*