College of Food Science and Light Industry, Nanjing Tech University , Nanjing, China.
Zhejiang Provincial Key Laboratory of Health Risk Factors for Seafood, Department of Food Science and Pharmacy, Zhejiang Ocean University , Zhoushan, China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2020 Jul;37(7):1061-1074. doi: 10.1080/19440049.2020.1746411. Epub 2020 May 12.
Dried squid products are popular in China as a snack food, side dishes, or refreshments, and the market appeal can be reflected by the high price that occasionally reaches 497 RMB per kg. However, the absence of harmonisation around the definition of squid, as well as the problems with visual inspection for processed seafood products, make alternative species substitution for dried squid products a frequent occurrence. The aim of the present study was to apply a DNA barcoding approach for species identification of 48 dried squid products collected from the largest online shopping platform in China. Moreover, we also developed a novel SYBR green real-time PCR assay (simplex and duplex followed by a melting curve analysis) specific for and based on cytochrome C oxidase subunit I (COI) gene. Results highlighted the successful DNA extraction and PCR amplification of a 655 bp COI gene fragment from all products. A maximum similarity value in the range of 98-100% was obtained for all readable sequences using the BOLD and BLAST public databases and four species (, and ) were identified. The specificity of the designed primer sets was confirmed against 23 non-target species, and the newly developed methods were successfully applied to screen and in dried squid products. Overall, DNA barcoding is a robust tool for seafood species identification and the novel method is effective in screening and in food products.
干鱿鱼产品作为中国的一种小吃、配菜或点心很受欢迎，其市场吸引力体现在其偶尔高达每公斤 497 元的高价上。然而，由于鱿鱼的定义没有统一，以及加工海产品的目视检查存在问题，因此经常会出现替代品种来冒充干鱿鱼产品。本研究旨在应用 DNA 条形码技术对从中国最大的在线购物平台上收集的 48 种干鱿鱼产品进行物种鉴定。此外，我们还根据细胞色素 C 氧化酶亚基 I (COI) 基因，开发了一种针对 和 的新型 SYBR 绿色实时 PCR 检测法（单重和双重反应，随后进行熔解曲线分析）。结果表明，所有产品均成功提取并扩增了 655 bp 的 COI 基因片段。使用 BOLD 和 BLAST 公共数据库对所有可读序列进行分析，得到的最大相似度值在 98-100%之间，鉴定出了 4 个物种（、、和）。对 23 种非目标物种进行了设计引物的特异性验证，新开发的方法成功应用于干鱿鱼产品中筛查 和 。总的来说，DNA 条形码是一种用于海鲜物种鉴定的强大工具，而新方法在筛选食品中的 和 方面非常有效。
