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用于三种梭子蟹物种快速鉴定的环介导等温扩增(LAMP)检测方法的开发

Development of Loop-Mediated Isothermal Amplification (LAMP) Assays for the Rapid Authentication of Three Swimming Crab Species.

作者信息

Benjakul Soottawat, Saetang Jirakrit

机构信息

International Center of Excellence in Seafood Science and Innovation, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai 90110, Songkhla, Thailand.

出版信息

Foods. 2022 Jul 28;11(15):2247. doi: 10.3390/foods11152247.

Abstract

Blue swimming crab meat is easily adulterated by other crab meats with a lower price. A potential authentication method is required to prevent mislabeling. LAMP assays were established to identify the meat of blue swimming crab, crucifix crab, and three spotted swimming crab. The primers were designed using PrimerExplorer V5. The specificity of the LAMP assay was tested compared to the PCR method. The sensitivity was conducted at the DNA concentrations of 0.4-50 ng/reaction. The results demonstrated that both LAMP and PCR could discriminate all species of crabs. LAMP showed a superior sensitivity to PCR in the three spotted swimming crab, while a similar result between LAMP and PCR was obtained in blue swimming crab. No changes in the detection efficacy were attained when boiled and steamed crab meats were applied. Therefore, the LAMP assay developed could potentially be applicable to detect the adulteration or mislabeling of raw or cooked crab meat in markets.

摘要

青蟹肉很容易被价格较低的其他蟹肉掺假。需要一种潜在的鉴定方法来防止标签错误。建立了环介导等温扩增(LAMP)检测方法来鉴定青蟹、十字蟹和三疣梭子蟹的肉。使用PrimerExplorer V5设计引物。与聚合酶链反应(PCR)方法相比,测试了LAMP检测的特异性。在DNA浓度为0.4 - 50 ng/反应时进行灵敏度检测。结果表明,LAMP和PCR都能区分所有蟹类物种。在三疣梭子蟹中,LAMP对PCR显示出更高的灵敏度,而在青蟹中LAMP和PCR得到了相似的结果。当应用煮熟和蒸熟的蟹肉时,检测效果没有变化。因此,所开发的LAMP检测方法有可能适用于检测市场上生的或熟的蟹肉掺假或标签错误。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b278/9368355/d4e84662eb0f/foods-11-02247-g001.jpg

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