Hubalkova Zora, Kralik Petr, Kasalova Janka, Rencova Eva
Department of Analytical Biotechnology and Department of Food and Feed Safety, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
J Agric Food Chem. 2008 May 28;56(10):3454-9. doi: 10.1021/jf703684w. Epub 2008 May 3.
Identification of fish species is significant due to the increasing interest of consumers in the meat of sea fish. Methods focusing on fish species identification help to reveal fraudulent substitution among economically important gadoid species in commercial seafood products. The objective of this work was to develop a conventional PCR method for the differentiation of the following gadoid fish species in fish products: Alaska pollack ( Theragra chalcogramma), blue whiting ( Micromesistius poutassou), hake spp. ( Merluccius spp.), Atlantic cod ( Gadus morhua), saithe ( Pollachius virens), and whiting ( Merlangius merlangus). The species-specific primer pairs for gadoid species determination were based on the partial pantophysin I ( PanI) genomic sequence. Sequence identification was confirmed by cloning and sequencing of the PCR products obtained from the species considered. For the simultaneous detection of Alaska pollack, blue whiting, and hake spp., a quadruplex PCR system was constructed. Other gadoid species were detected in separate PCR reactions. After optimization of the reactions, the developed PCR systems were used for the analysis of codfish samples obtained from the Czech market and the customs' laboratories. This method represents an alternative approach in the use of genomic DNA for the identification of fish species. This method is rapid, simple, and reliable without the need for further confirmative methods. Furthermore, the identification of a mixture of more than one species is possible. The PCR system has been optimized for routine diagnostic purposes.
由于消费者对海鱼鱼肉的兴趣日益增加，鱼类物种鉴定具有重要意义。专注于鱼类物种鉴定的方法有助于揭示商业海产品中经济上重要的鳕形目鱼类之间的欺诈性替代。这项工作的目的是开发一种常规PCR方法，用于区分鱼类产品中的以下鳕形目鱼类物种：阿拉斯加狭鳕鱼（黄线狭鳕）、蓝鳕（小鳍鳕）、无须鳕属（多种无须鳕）、大西洋鳕鱼（大西洋鳕）、黑线鳕（绿青鳕）和牙鳕（欧洲牙鳕）。用于确定鳕形目物种的物种特异性引物对基于部分泛素蛋白I（PanI）基因组序列。通过对从所考虑的物种获得的PCR产物进行克隆和测序来确认序列鉴定。为了同时检测阿拉斯加狭鳕鱼、蓝鳕和无须鳕属，构建了四重PCR系统。其他鳕形目物种在单独的PCR反应中进行检测。在对反应进行优化后，所开发的PCR系统用于分析从捷克市场和海关实验室获得的鳕鱼样本。这种方法代表了一种使用基因组DNA鉴定鱼类物种的替代方法。该方法快速、简单且可靠，无需进一步的确认方法。此外，还可以鉴定一种以上物种的混合物。该PCR系统已针对常规诊断目的进行了优化。
