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利用线粒体 DNA 序列的小沟结合探针实时聚合酶链反应分析鉴定商业罐装金枪鱼品种。

Identification of tuna species in commercial cans by minor groove binder probe real-time polymerase chain reaction analysis of mitochondrial DNA sequences.

机构信息

Department of Veterinary Public Health, Faculty of Veterinary Medicine of Bari, Strada per Casamassima km 3, 70010 Valenzano, Bari, Italy.

出版信息

Mol Cell Probes. 2010 Dec;24(6):352-6. doi: 10.1016/j.mcp.2010.07.006. Epub 2010 Aug 4.

Abstract

Three different minor groove binder (MGB) probe assays have been developed for rapid and accurate identification of the species commonly used for production of canned tuna, i.e. yellowfin (Thunnus albacares), bluefin (Thunnus thynnus) and albacore (Thunnus alalunga) tunas. The assays targeting the mitochondrial cytochrome b gene were able to discriminate efficiently between the three species contained in fresh or canned tunas and did not react with other Scombroidei that were tested. A correct species prediction was obtained even from artificial mixtures prepared with different amounts of the reference tuna species and subjected to the sterilisation treatment. Testing of 27 commercial canned tunas by PCR-RFLP, MGB probe assays and sequence analysis showed a concordance of 100% between the last two techniques, whereas by using PCR-RFLP several samples were uncharacterised or mischaracterised. These results make the established MGB probe assays an attractive tool for direct and rapid species identification in canned tuna.

摘要

三种不同的小沟结合物(MGB)探针检测方法已被开发出来,用于快速准确地鉴定用于生产罐头金枪鱼的常见物种,即黄鳍金枪鱼(Thunnus albacares)、蓝鳍金枪鱼(Thunnus thynnus)和长鳍金枪鱼(Thunnus alalunga)。针对线粒体细胞色素 b 基因的检测方法能够有效地区分新鲜或罐装金枪鱼中的这三个物种,并且不会与其他测试的鲭鱼科鱼类发生反应。即使是用不同数量的参考金枪鱼物种制备并经过灭菌处理的人工混合物,也能得到正确的物种预测。通过 PCR-RFLP、MGB 探针检测和序列分析对 27 个商业罐头金枪鱼进行测试,结果显示后两种技术之间的一致性为 100%,而使用 PCR-RFLP 时,一些样本无法被识别或被错误识别。这些结果使建立的 MGB 探针检测方法成为一种有吸引力的工具,可用于直接快速鉴定罐头金枪鱼的物种。

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