Chapela Maria Jose, Sánchez Ana, Suárez Maria Isabel, Pérez-Martín Ricardo I, Sotelo Carmen G
Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas, Eduardo Cabello 6, Vigo 36208 Pontevedra, Spain.
J Agric Food Chem. 2007 Aug 22;55(17):6903-9. doi: 10.1021/jf070132c. Epub 2007 Jul 24.
An accurate screening method for hake species identification based in single-stranded conformation polymorphism analysis is presented. The differentiation of 11 species of the Merluccius genus and another five species of the Gadiformes order was studied. For this purpose, two fragments of the cytochrome b gene were sequenced; the first is the 5'-end, a fragment of 465 bp (Kocher fragment), and the second is the 3'-end of the cytochrome b, a 588 bp fragment (SB fragment). These two fragments were amplified, denatured, and submitted to native nondenaturing polyacrylamide gel electrophoresis. Results show that with this technique and both fragments, all of the species studied can be unequivocally identified. The validation of the methodology was carried out with 24 commercial hake products showing good performance of the technique for species identification in commercial products. Results show that all species were identified. This technique has advantages over other published methods, because only one polymerase chain reaction step is needed, saving time and money, and it decreases the time needed for hake species identification in food products, making it especially suitable as a screening methodology when a high number of samples should be analyzed in routine examinations.
本文介绍了一种基于单链构象多态性分析的无须鳕物种鉴定的精确筛选方法。研究了无须鳕属11个物种以及鳕形目另外5个物种的鉴别。为此，对细胞色素b基因的两个片段进行了测序；第一个是5'端，一个465 bp的片段（科赫尔片段），第二个是细胞色素b的3'端，一个588 bp的片段（SB片段）。这两个片段经过扩增、变性，然后进行非变性聚丙烯酰胺凝胶电泳。结果表明，使用该技术和这两个片段，可以明确鉴定所有研究的物种。该方法通过对24种商业无须鳕产品进行验证，结果表明该技术在商业产品的物种鉴定中表现良好。结果显示所有物种均被鉴定出来。该技术相对于其他已发表的方法具有优势，因为只需要一个聚合酶链反应步骤，节省了时间和金钱，并且减少了食品中无须鳕物种鉴定所需的时间，使其特别适合作为一种筛选方法，用于在常规检查中分析大量样本。
