Ye Jian, Feng Junli, Liu Shasha, Zhang Yanping, Jiang Xiaona, Dai Zhiyuan
Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China.
Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou, Zhejiang 310018, PR China.
Mol Cell Probes. 2016 Feb;30(1):22-9. doi: 10.1016/j.mcp.2016.01.001. Epub 2016 Jan 7.
Squids are distributed worldwide, including many species of commercial importance, and they are often made into varieties of flavor foods. The rapid identification methods for squid species especially their processed products, however, have not been well developed. In this study, quantitative real-time PCR (qPCR) systems based on specific primers and TaqMan probes have been established for rapid and accurate identification of four common squid species (Ommastrephes bartramii, Dosidicus gigas, Illex argentinus, Todarodes pacificus) in Chinese domestic market. After analyzing mitochondrial genes reported in GenBank, the mitochondrial cytochrome b (Cytb) gene was selected for O. bartramii detection, cytochrome c oxidase subunit I (COI) gene for D. gigas and T. Pacificus detection, ATPase subunit 6 (ATPase 6) gene for I. Argentinus detection, and 12S ribosomal RNA (12S rDNA) gene for designing Ommastrephidae-specific primers and probe. As a result, all the TaqMan systems are of good performance, and efficiency of each reaction was calculated by making standard curves. This method could detect target species either in single or mixed squid specimen, and it was applied to identify 12 squid processed products successfully. Thus, it would play an important role in fulfilling labeling regulations and squid fishery control.
鱿鱼分布于世界各地，包括许多具有商业重要性的种类，并且它们常被制成各种风味食品。然而，鱿鱼种类尤其是其加工产品的快速鉴定方法尚未得到很好的发展。在本研究中，基于特异性引物和TaqMan探针建立了定量实时PCR（qPCR）系统，用于快速准确地鉴定中国国内市场上的四种常见鱿鱼种类（柔鱼、茎柔鱼、阿根廷滑柔鱼、太平洋褶柔鱼）。在分析GenBank中报道的线粒体基因后，选择线粒体细胞色素b（Cytb）基因用于检测柔鱼，细胞色素c氧化酶亚基I（COI）基因用于检测茎柔鱼和太平洋褶柔鱼，ATP酶亚基6（ATPase 6）基因用于检测阿根廷滑柔鱼，12S核糖体RNA（12S rDNA）基因用于设计柔鱼科特异性引物和探针。结果，所有TaqMan系统性能良好，并通过制作标准曲线计算每个反应的效率。该方法可以在单一或混合鱿鱼样本中检测目标种类，并成功应用于鉴定12种鱿鱼加工产品。因此，它将在履行标签规定和鱿鱼渔业控制方面发挥重要作用。
