title: Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood.
abstract: A real time PCR assay for the detection of Vibrio parahaemolyticus in seafood samples was developed using a novel specific target and a competitive internal amplification control (IAC). The specificity of this assay was evaluated using 390 bacterial strains including V. parahaemolyticus, and other strains belonging to Vibrio and non-Vibrio species. The real time PCR assay unambiguously distinguished V. parahaemolyticus with a detection sensitivity of 4.8 fg per PCR with purified genomic DNA or 1 CFU per reaction by counting V. parahaemolyticus colonies. The assays of avoiding interference demonstrated that, even in the presence of 2.1 μg genomic DNA or 10(7) CFU background bacteria, V. parahaemolyticus could still be accurately detected. In addition, the IAC was used to indicate false-negative results, and lower than 94 copies of IAC per reaction had no influence on the detection limit. Ninety-six seafood samples were tested, of which 58 (60.4%) were positive, including 3 false negative results. Consequently, the real time PCR assay is effective for the rapid detection of V. parahaemotyticus contaminants in seafood.
authors.affiliation: MOST-USDA Joint Research Center for Food Safety and Bor Luh Food Safety Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.forename: Xiaohua
authors.initials: X
authors.lastname: He
authors.forename: Wanyi
authors.initials: W
authors.lastname: Chen
authors.forename: Shuijing
authors.initials: S
authors.lastname: Yu
authors.forename: Chunlei
authors.initials: C
authors.lastname: Shi
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Zhou
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Dapeng
authors.initials: D
authors.lastname: Wang
authors.forename: Xianming
authors.initials: X
authors.lastname: Shi
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry / Q000378:metabolism; D005516:Food Microbiology / Q000379:methods*; D016680:Genome, Bacterial; D060888:Real-Time Polymerase Chain Reaction; D017747:Seafood / Q000382:microbiology*; D014733:Vibrio / Q000235:genetics / Q000302:isolation & purification; D014736:Vibrio parahaemolyticus / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D018528:ATP-Binding Cassette Transporters; D017931:DNA Primers