title: Development of a qPCR Method for the Identification and Quantification of Two Closely Related Tuna Species, Bigeye Tuna (Thunnus obesus) and Yellowfin Tuna (Thunnus albacares), in Canned Tuna.
abstract: Bigeye tuna (Thunnus obesus) and yellowfin tuna (Thunnus albacares) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, as juveniles of bigeye and yellowfin tunas are very difficult to distinguish, unintentional substitutions may occur during the canning process. In this study, two mitochondrial markers from NADH dehydrogenase subunit 2 and cytochrome c oxidase subunit II genes were used to identify bigeye tuna and yellowfin tuna, respectively, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one was based on absolute quantification using standard curves realized with these two markers; the second one was founded on relative quantification with the universal 12S rRNA gene as the endogenous gene. On the basis of our results, we conclude that our methodology could be applied to authenticate these two closely related tuna species when used in a binary mix in tuna cans.
authors.affiliation: Université Littoral Côte d'Opale , EA 7394 - ICV - Institut Charles Viollette, USC Anses - ULCO, F-62200 Boulogne-sur-Mer, France.|Laboratoire d'Océanologie et de Géosciences, UMR 8187 (ULCO, Lille 1, CNRS) , 62930 Wimereux, France.|PFINV , F-62200 Boulogne-sur-Mer, France.
authors.forename: Daline
authors.initials: D
authors.lastname: Bojolly
authors.affiliation: Université Littoral Côte d'Opale , EA 7394 - ICV - Institut Charles Viollette, USC Anses - ULCO, F-62200 Boulogne-sur-Mer, France.|Université Lille , F-59000 Lille, France.|Université Artois , F-62000 Arras, France.|INRA , France.|ISA , F-59000 Lille, France.
authors.forename: Périne
authors.initials: P
authors.lastname: Doyen
authors.affiliation: PFINV , F-62200 Boulogne-sur-Mer, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Le Fur
authors.affiliation: Laboratoire d'Océanologie et de Géosciences, UMR 8187 (ULCO, Lille 1, CNRS) , 62930 Wimereux, France.
authors.forename: Urania
authors.initials: U
authors.lastname: Christaki
authors.affiliation: Ifremer , F-44300 Nantes, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Verrez-Bagnis
authors.affiliation: Université Littoral Côte d'Opale , EA 7394 - ICV - Institut Charles Viollette, USC Anses - ULCO, F-62200 Boulogne-sur-Mer, France.|Université Lille , F-59000 Lille, France.|Université Artois , F-62000 Arras, France.|INRA , France.|ISA , F-59000 Lille, France.
authors.forename: Thierry
authors.identifier: http://orcid.org/0000-0002-4636-8981
authors.initials: T
authors.lastname: Grard
mesh_terms: D000818:Animals; D016002:Discriminant Analysis; D003576:Electron Transport Complex IV / Q000235:genetics; D005396:Fish Products / Q000032:analysis*; D029941:Fish Proteins / Q000235:genetics; D005506:Food Contamination / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014413:Tuna / Q000145:classification / Q000235:genetics*
chemical_list: D029941:Fish Proteins; C117108:cytochrome C oxidase subunit II; D003576:Electron Transport Complex IV
keywords: TaqMan; authentication; bigeye tuna (Thunnus obesus); canned products; qPCR; quantification; species identification; tuna; yellowfin tuna (Thunnus albacares)