title: Monitoring the freshness of fish: development of a qPCR method applied to MAP chilled whiting.
abstract: BACKGROUND
  Monitoring of early stages of freshness decay is a major issue for the fishery industry to guarantee the best quality for this highly perishable food matrix. Numerous techniques have been developed, but most of them have the disadvantage of being reliable only either in the last stages of fish freshness or for the analysis of whole fish. This study describes the development of a qPCR method targeting the torA gene harboured by fish spoilage microorganisms. torA encodes an enzyme that leads to the production of trimethylamine responsible for the characteristic spoiled-fish odour.
  
  
  RESULTS
  A degenerate primer pair was designed. It amplified torA gene of both Vibrio and Photobacterium with good efficiencies on 7-log DNA dilutions. The primer pair was used during a shelf-life monitoring study achieved on modified atmosphere packed, chilled, whiting (Merlangius merlangus) fillets. The qPCR approach allows the detection of an increase of torA copies throughout the storage of fillets in correlation with the evolution of both total volatile basic nitrogen (-0.86) and trimethylamine concentrations (-0.81), known as spoilage markers.
  
  
  CONCLUSIONS
  This study described a very promising, sensitive, reliable, time-effective, technique in the field of freshness characterisation of processed fish.
authors.affiliation: ANSES, Laboratoire de Sécurité des Aliments - Département des Produits de la Pêche et de l'Aquaculture, Boulevard du Bassin Napoléon, 62200, Boulogne-sur-Mer, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Dehaut
authors.affiliation: Université de Lille 1, Unité de Glycobiologie Structurale et Fonctionnelle - UGSF - Bâtiment C9, 59650, Villeneuve d'Ascq, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Krzewinski
authors.affiliation: Université du Littoral Côte d'Opale - ANSES, Équipe Biochimie des Produits Aquatiques, Boulevard du Bassin Napoléon, 62200, Boulogne-sur-mer, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Grard
authors.affiliation: Laboratoire Régional de Recherche en Agroalimentaire et Biotechnologie - Institut Charles Viollette, Equipe ProBioGEM, Université de Lille1, Sciences et Technologies, Polytech'Lille, Avenue Paul Langevin, 59655, Villeneuve d'Ascq, France.
authors.forename: Marlène
authors.initials: M
authors.lastname: Chollet
authors.affiliation: Laboratoire Régional de Recherche en Agroalimentaire et Biotechnologie - Institut Charles Viollette, Equipe ProBioGEM, Université de Lille1, Sciences et Technologies, Polytech'Lille, Avenue Paul Langevin, 59655, Villeneuve d'Ascq, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Jacques
authors.affiliation: ANSES, Laboratoire de Sécurité des Aliments - Département des Produits de la Pêche et de l'Aquaculture, Boulevard du Bassin Napoléon, 62200, Boulogne-sur-Mer, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Brisabois
authors.affiliation: ANSES, Laboratoire de Sécurité des Aliments - Département des Produits de la Pêche et de l'Aquaculture, Boulevard du Bassin Napoléon, 62200, Boulogne-sur-Mer, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Duflos
mesh_terms: D000818:Animals; D001272:Atmosphere; D003080:Cold Temperature; D029941:Fish Proteins / Q000737:chemistry; D005399:Fishes; D018857:Food Packaging / Q000379:methods*; D063427:Food Quality; D008460:Meat / Q000592:standards*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D029941:Fish Proteins
keywords: freshness assessment; modified atmosphere packed fish; qPCR; torA