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中国精品科技期刊2020
彭博,洪麟,高世光,等. 食品中三文鱼源性成分等温快检方法建立J. 食品工业科技,2026,47(4):1−9. doi: 10.13386/j.issn1002-0306.2025030257.
引用本文: 彭博,洪麟,高世光,等. 食品中三文鱼源性成分等温快检方法建立J. 食品工业科技,2026,47(4):1−9. doi: 10.13386/j.issn1002-0306.2025030257.
PENG Bo, HONG Lin, GAO Shiguang, et al. Establishment of Isothermal Rapid Detection Method for Salmon-derived Components in FoodJ. Science and Technology of Food Industry, 2026, 47(4): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2025030257.
Citation: PENG Bo, HONG Lin, GAO Shiguang, et al. Establishment of Isothermal Rapid Detection Method for Salmon-derived Components in FoodJ. Science and Technology of Food Industry, 2026, 47(4): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2025030257.

食品中三文鱼源性成分等温快检方法建立

Establishment of Isothermal Rapid Detection Method for Salmon-derived Components in Food

  • 摘要: 本文建立了虹鳟和大西洋鲑两种三文鱼源性成分可视化和荧光环介导等温扩增(Loop-mediated isothermal amplification,LAMP)及重组酶聚合酶扩增(Recombinase polymerase amplification, RPA)快速检测方法。以COI基因作为靶标基因,设计筛选适于等温扩增的引物序列,开展等温扩增方法的特异性和灵敏度分析,同时比较基于快速DNA提取和磁珠提取DNA的LAMP方法的适用性。结果表明,所构建的LAMP和RPA等温扩增方法特异性强,使用磁珠法提取DNA的LAMP检测的灵敏度分别可达到2.99×10−1 ng/μL和5.16×10−3 ng/μL,使用快速提取DNA的LAMP检测灵敏度分别可达到9.42×100 ng/μL和1.13×101 ng/μL。同时制备模拟混合样品,发现虹鳟和大西洋鲑成分RPA等温扩增的灵敏度可达到0.1% (W/W),可视化LAMP等温扩增的灵敏度可稳定达到0.5% (W/W)。LAMP和RPA方法满足低技术要求、低设备成本以及检测时间短的快速检测要求。同时使用裂解液快速提取基因组DNA,可以在30~40 min之内完成检测,对于生产加工及市场等流通环节现场进行三文鱼成分真实性检测是一个很好的选择。

     

    Abstract: This article presented a visualization method for detecting salmon-derived components in Oncorhynchus mykiss and Salmo salar, based on fluorescence-labeled loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA). The widely recognized COI gene was targeted to design and screen optimal primer sequences for isothermal amplification. The specificity and sensitivity of the isothermal amplification methods were analyzed, and the applicability of LAMP using rapid DNA extraction and magnetic bead-based DNA extraction techniques was compared. The results demonstrated that the constructed LAMP and RPA methods exhibited high specificity. The sensitivity of LAMP detection using magnetic bead-extracted DNA reached 2.99×10−1 ng/μL and 5.16×10−3 ng/μL, respectively, whereas using rapid DNA extraction, sensitivity reached 9.42×100 ng/μL and 1.13×101 ng/μL, respectively. Simulated mixed samples were prepared, and results showed that the sensitivity of RPA amplification for detecting Oncorhynchus mykiss and Salmo salar components reached 0.1% (w/w), while the sensitivity of visual LAMP amplification consistently reached 0.5% (w/w). Both LAMP and RPA methods met rapid detection requirements, characterized by low technical complexity, minimal equipment cost, and short detection time. Moreover, rapid DNA extraction using lysis solutions allowed the detection process to be completed within 30~40 min, making this approach an excellent choice for on-site authenticity testing of fish species in production, processing, and market circulation stages.

     

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