title: Development of a real-time PCR method for the identification of Atlantic mackerel (Scomber scombrus).
abstract: A Real Time-PCR method based on TaqMan technology for the identification of Scomber scombrus has been developed. A system of specific primers and a Minor Groove Binding (MGB) TaqMan probe based on sequences of the mitochondrial cytochrome b region was designed. The method was successfully tested in 81 specimens of S. scombrus and related species and validated in 26 different commercial samples. An average Threshold Cycle (Ct) value of 15.3 was obtained with S. scombrus DNA. With the other species tested fluorescence signal was not detected or Ct was significantly higher (P<0.001). The efficiency of the assay was estimated to be 92.41%, with 100% specificity, and no cross reactivity was detected with any other species. These results reveal that the developed method is a rapid and efficient tool to unequivocally identify S. scombrus and may aid in the prevention of fraud or mislabelling in mackerel products.
authors.affiliation: Instituto de Investigaciones Marinas (IIM-CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain. amayavelasco@iim.csic.es
authors.forename: Amaya
authors.initials: A
authors.lastname: Velasco
authors.forename: Ana
authors.initials: A
authors.lastname: Sánchez
authors.forename: Icíar
authors.initials: I
authors.lastname: Martínez
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Santaclara
authors.forename: Ricardo I
authors.initials: RI
authors.lastname: Pérez-Martín
authors.forename: Carmen G
authors.initials: CG
authors.lastname: Sotelo
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D029941:Fish Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D010473:Perciformes / Q000145:classification / Q000235:genetics*; D011786:Quality Control; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017747:Seafood / Q000032:analysis*
chemical_list: D017931:DNA Primers; D029941:Fish Proteins
keywords: Atlantic mackerel; Cytochrome b; Pelagic fish; Real-Time PCR; Scomber; Species identification