Développement d'une méthode méthodologie de PCR en temps réel pour l'identification et la quantification de trois espèces de thon (Thunnus obesus, Thunnus albacares et Katsuwonus pelamis) dans les produits appertisés

Abstract : Bigeye tuna (Thunnus obesus), yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelanis) 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, it can be difficult to authenticate the tuna species, due to their high degree of similarity or even when the external morphological characteristics are removed due to filleting before canning. Consequently, involuntary or fraudulent substitutions may occur during the canning process. In this study, the mitochondrial marker from NADH dehydrogenase subunit 2 gene was used to identify bigeye tuna and the mitochondrial marker cytochrome c oxidase subunit II gene was used to identify yellowfin tuna and skipjack tuna, 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 the two closely related tuna species (bigeye tuna and yellowfin tuna) when used in a binary mix in tuna cans.
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Daline Bojolly. Développement d'une méthode méthodologie de PCR en temps réel pour l'identification et la quantification de trois espèces de thon (Thunnus obesus, Thunnus albacares et Katsuwonus pelamis) dans les produits appertisés. Sciences et techniques des pêches. Université du Littoral Côte d'Opale, 2017. Français. ⟨NNT : 2017DUNK0445⟩. ⟨tel-02151058⟩

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