Bottero Maria Teresa, Dalmasso Alessandra, Cappelletti Marco, Secchi Camillo, Civera Tiziana
Department of Animal Pathology, Faculty of Veterinary Medicine, University of Torino, via Leonardo da Vinci 44, 10095 Grugliasco, Torino, Italy.
J Biotechnol. 2007 May 1;129(3):575-80. doi: 10.1016/j.jbiotec.2007.01.032. Epub 2007 Feb 11.
A novel methodology based on analysis of mtDNA-cytb diagnostic sites was performed to discriminate four closely related species of Thunnus (Thunnus alalunga, Thunnus albacares, Thunnus obesus and Thunnus thynnus) and one species of Euthynnus (Katsuwonus pelamis) genus in raw and canned tuna. The primers used in the preliminary PCR designed in well conserved region upstream and downstream of the diagnosis sites successfully amplified a 132bp region from the cytb gene of all the species taken into consideration. The sites of diagnosis have been interrogate simultaneously using a multiplex primer-extension assay (PER) and the results were confirmed by fragment sequencing. The applicability of the multiplex PER assay to commercial canned tuna samples was also demonstrated. The proposed test could be useful for detection of fraud and for seafood traceability.
基于线粒体DNA细胞色素b（mtDNA-cytb）诊断位点分析的一种新方法，用于鉴别生的和罐装金枪鱼中鲔属的四种近缘物种（大西洋蓝鳍金枪鱼、黄鳍金枪鱼、大眼金枪鱼和蓝鳍金枪鱼）以及狐鲣属的一种物种（鲣鱼）。在诊断位点上下游高度保守区域设计的初步聚合酶链式反应（PCR）中使用的引物，成功地从所有考虑的物种的细胞色素b基因中扩增出一个132bp的区域。使用多重引物延伸分析（PER）同时检测诊断位点，并通过片段测序确认结果。还证明了多重PER分析在商业罐装金枪鱼样品中的适用性。所提出的检测方法对于欺诈检测和海鲜可追溯性可能是有用的。
