Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna

Tuna are economically important as food resources in food markets. However, because tuna is often processed into steaks or fillets, the meat can be difficult to identify through morphological features. For effective fishery management and to protect the rights of consumers, it is necessary to develo...

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Main Authors: Meng Qu, Yanhua Jiang, Na Li, Yingying Guo, Wenjia Zhu, Xinnan Zhao, Lin Yao, Lianzhu Wang
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/13/22/3692
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author Meng Qu
Yanhua Jiang
Na Li
Yingying Guo
Wenjia Zhu
Na Li
Xinnan Zhao
Lin Yao
Lianzhu Wang
author_facet Meng Qu
Yanhua Jiang
Na Li
Yingying Guo
Wenjia Zhu
Na Li
Xinnan Zhao
Lin Yao
Lianzhu Wang
author_sort Meng Qu
collection DOAJ
description Tuna are economically important as food resources in food markets. However, because tuna is often processed into steaks or fillets, the meat can be difficult to identify through morphological features. For effective fishery management and to protect the rights of consumers, it is necessary to develop a molecular method to accurately identify the species used in tuna products. Herein, we discovered five single-nucleotide polymorphism (SNP) sites via 2b-RAD sequencing and developed five SNP-based real-time polymerase chain reaction assays for the rapid identification of five highly priced tuna species. Three species-specific TaqMan systems were designed to identify albacore tuna (<i>Thunnus alalunga</i>), bigeye tuna (<i>T. obesus</i>), and southern bluefin tuna (<i>T. maccoyii</i>) and two cycling systems were designed to identify yellowfin tuna (<i>T. albacares</i>) and Atlantic bluefin tuna (<i>T. thynnus</i>). The systems showed good specificity and sensitivity (sensitivity of 0.0002 ng μL<sup>−1</sup> for albacore tuna, bigeye tuna, and southern bluefin tuna and 0.002 ng μL<sup>−1</sup> for yellowfin tuna and Atlantic bluefin tuna). Both systems were able to distinguish the target species from other species in a specific, sensitive, and accurate manner. Thus, these methods can be employed for the identification of species used in tuna products, protecting consumers and producers from economic fraud.
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spelling doaj-art-cbb1d2a36302424dbc7ac3fa81c989b82025-08-20T02:28:10ZengMDPI AGFoods2304-81582024-11-011322369210.3390/foods13223692Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced TunaMeng Qu0Yanhua Jiang1Na Li2Yingying Guo3Wenjia Zhu4Na Li5Xinnan Zhao6Lin Yao7Lianzhu Wang8Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaShandong Institute for Product Quality Inspection, Jinan 250102, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaKey Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaTuna are economically important as food resources in food markets. However, because tuna is often processed into steaks or fillets, the meat can be difficult to identify through morphological features. For effective fishery management and to protect the rights of consumers, it is necessary to develop a molecular method to accurately identify the species used in tuna products. Herein, we discovered five single-nucleotide polymorphism (SNP) sites via 2b-RAD sequencing and developed five SNP-based real-time polymerase chain reaction assays for the rapid identification of five highly priced tuna species. Three species-specific TaqMan systems were designed to identify albacore tuna (<i>Thunnus alalunga</i>), bigeye tuna (<i>T. obesus</i>), and southern bluefin tuna (<i>T. maccoyii</i>) and two cycling systems were designed to identify yellowfin tuna (<i>T. albacares</i>) and Atlantic bluefin tuna (<i>T. thynnus</i>). The systems showed good specificity and sensitivity (sensitivity of 0.0002 ng μL<sup>−1</sup> for albacore tuna, bigeye tuna, and southern bluefin tuna and 0.002 ng μL<sup>−1</sup> for yellowfin tuna and Atlantic bluefin tuna). Both systems were able to distinguish the target species from other species in a specific, sensitive, and accurate manner. Thus, these methods can be employed for the identification of species used in tuna products, protecting consumers and producers from economic fraud.https://www.mdpi.com/2304-8158/13/22/3692tunaspecies identification2b-RADSNPreal-time PCR
spellingShingle Meng Qu
Yanhua Jiang
Na Li
Yingying Guo
Wenjia Zhu
Na Li
Xinnan Zhao
Lin Yao
Lianzhu Wang
Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
Foods
tuna
species identification
2b-RAD
SNP
real-time PCR
title Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
title_full Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
title_fullStr Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
title_full_unstemmed Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
title_short Development of Single-Nucleotide Polymorphism (SNP)-Based Species-Specific Real-Time PCR Assays for Authenticating Five Highly Priced Tuna
title_sort development of single nucleotide polymorphism snp based species specific real time pcr assays for authenticating five highly priced tuna
topic tuna
species identification
2b-RAD
SNP
real-time PCR
url https://www.mdpi.com/2304-8158/13/22/3692
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