Simple PCR-RFLP detection method for genus- and species-authentication of four types of tuna used in canned tuna industry

© 2019 Elsevier Ltd A simple method for authentication of processed tuna products is not only for testing against fraudulent practices in the tuna industry but being a promising tool for enhancing traceability. Here, a method of polymerase chain reaction-restriction fragment length polymorphism (PCR...

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Bibliographic Details
Main Authors: Wanniwat Mata, Thanakorn Chanmalee, Napassorn Punyasuk, Siripong Thitamadee
Other Authors: Mahidol University
Format: Article
Published: 2020
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/49511
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Institution: Mahidol University
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Summary:© 2019 Elsevier Ltd A simple method for authentication of processed tuna products is not only for testing against fraudulent practices in the tuna industry but being a promising tool for enhancing traceability. Here, a method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) had been developed using a part of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene as a key region for genus-and species-identification. The analysis was based on the different RFLP patterns of TaqI and HaeIII digested COI DNA from the four tuna species; i.e. Kasuwonus pelamis, Thunnus alalunga, Thunnus albacares, and Thunnus obesus, respectively. While the conventional PCR-RFLP worked effectively in raw tuna meat, a method of semi-nested PCR had to be developed in order to increase efficiency in the detection in cooked meat from canned product. The semi-nested PCR-RFLP pattern of COI DNA allowed the detection of 10% contamination of K. pelamis in canned tuna of other higher-valued species.