重组酶聚合酶扩增结合侧流试纸快速鉴定大西洋鳕鱼、貂鱼和犬牙鱼：在中国鳕鱼相关鱼类产品检测中的应用

IF 5.6 1区农林科学 Q1 FOOD SCIENCE & TECHNOLOGY

Food Control Pub Date : 2025-04-11 DOI:10.1016/j.foodcont.2025.111356

Fangjie Chen , Zhongjie Xu , Yihuan Chen , Le Zhang , Shilin Shang , Liyuan Yi , Wenjie Wang , Zengyi Zhang , Qianjin Zhou , Jiong Chen

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引用次数: 0

摘要

Gadus因其优越的营养品质而受到高度重视；然而，频繁的错误标签引起了消费者的担忧。在中国，出于商业原因，Sablefish （Anoplopoma fibria）和toothfish （Dissostichus eleginoides和D. mawsoni）经常以包括“鳕鱼”在内的名称销售。本研究开发了一种重组酶聚合酶扩增法（RPA），结合横向流量试纸（LFD）快速检测大西洋鳕鱼（Gadus morhua）、貂鱼和齿鱼。为每个物种设计了两个特异性引物（包括一个生物素标记的反向引物）和一个带有5 ' FAM标记和thf取代胸腺嘧啶的探针，靶向细胞色素c氧化酶亚基I （COI）基因的种间区域。所开发的RPA-LFD检测方法对大西洋鳕鱼、貂鱼和犬牙鱼具有特异性，与其他硬骨鱼物种无交叉反应。重组质粒DNA检测限为1 ~ 100 copies/μL，重组质粒DNA检测限为1 ~ 100 fg/μL，重组质粒DNA检测限为大西洋鳕鱼、貂鱼和巴塔哥尼亚犬牙鱼。引物浓度均匀优化为0.4 μM，大西洋鳕鱼和巴塔哥尼亚犬牙鱼的探针浓度分别为0.12 μM和0.18 μM。RPA条件略有不同，每个物种的最佳持续时间为14-16分钟，温度为36-39℃。对来自中国大型零售商店的67种冷冻鳕鱼相关鱼类产品的检测显示，COI测序结果100%一致。结果表明，这些产品的误标率高达17.91%，但被替代的品种相对较少，主要有太平洋鳕鱼（M. poutassou）、阿拉斯加鳕鱼（G. chalcogrammus）和蓝鳕（Micromesistius poutassou）。RPA-LFD方法具有快速、简便、直观、灵敏、准确的特点，可用于大西洋鳕鱼、貂鱼和齿鱼的鉴定，可用于鳕鱼相关鱼类的现场掺假监测。

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Rapid identification of Atlantic cod, sablefish, and toothfish using recombinase polymerase amplification combined with lateral flow dipstick: Application in the detection of cod-related fish products in China

Gadus species are highly valued for their superior nutritional quality; however, frequent mislabeling raises consumer concerns. Sablefish (Anoplopoma fimbria) and toothfish (Dissostichus eleginoides and D. mawsoni) are often marketed in China under names that include “cod” for commercial reasons. This study developed a recombinase polymerase amplification assay (RPA) combined with a lateral flow dipstick (LFD) to rapidly detect Atlantic cod (Gadus morhua), sablefish, and toothfish. Two specific primers (including one biotin-labeled reverse primer) and a probe with 5′ FAM label and THF-substituted thymine were designed for each species, targeting the interspecific regions of the cytochrome c oxidase subunit I (COI) gene. The developed RPA-LFD assays were specific for Atlantic cod, sablefish, and toothfish, showing no cross-reactivity with other teleost species. The assays demonstrated detection limits of 1–100 copies/μL for recombinant plasmid DNA and 1–100 fg/μL for genomic DNA across Atlantic cod, sablefish, and Patagonian toothfish. Primer concentrations were uniformly optimized to 0.4 μM, while probe concentrations were set at 0.12 μM for Atlantic cod and Patagonian toothfish, and 0.18 μM for sablefish. RPA conditions varied slightly, with optimal durations of 14–16 min and temperatures of 36–39 °C tailored to each species. Testing of 67 frozen cod-related fish products from large retail stores in China showed 100 % concordance with COI sequencing. Additionally, the results revealed a high mislabeling rate of 17.91 % among these products, although the substituted species were relatively limited, primarily including Pacific cod (M. poutassou), Alaska pollock (G. chalcogrammus), and blue whiting (Micromesistius poutassou). This RPA-LFD assay is rapid, simple, visual, sensitive, and accurate for identifying Atlantic cod, sablefish, and toothfish, and could help monitor cod-related fish adulteration on site.

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来源期刊

Food Control 工程技术-食品科技

CiteScore

12.20

自引率

6.70%

发文量

758

审稿时长

33 days

期刊介绍： Food Control is an international journal that provides essential information for those involved in food safety and process control. Food Control covers the below areas that relate to food process control or to food safety of human foods: • Microbial food safety and antimicrobial systems • Mycotoxins • Hazard analysis, HACCP and food safety objectives • Risk assessment, including microbial and chemical hazards • Quality assurance • Good manufacturing practices • Food process systems design and control • Food Packaging technology and materials in contact with foods • Rapid methods of analysis and detection, including sensor technology • Codes of practice, legislation and international harmonization • Consumer issues • Education, training and research needs. The scope of Food Control is comprehensive and includes original research papers, authoritative reviews, short communications, comment articles that report on new developments in food control, and position papers.