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一种用于快速、特异性检测蓝鳕(Molva molva)的新型实时聚合酶链反应方法。

A new real-time PCR method for rapid and specific detection of ling (Molva molva).

作者信息

Taboada Ledicia, Sánchez Ana, Sotelo Carmen G

机构信息

Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain.

出版信息

Food Chem. 2017 Aug 1;228:469-475. doi: 10.1016/j.foodchem.2017.01.117. Epub 2017 Feb 8.

Abstract

Seafood fraud - often involving substitution of one species by another - has attracted much attention as it is prevalent worldwide. Whilst DNA analysis has helped to combat this type of fraud some of the methods currently in use are time-consuming and require sophisticated equipment or highly-trained personnel. This work describes the development of a new, real-time PCR TaqMan assay for the detection of ling (Molva molva) in seafood products. For this purpose, specific primers and a minor groove binding (MGB) TaqMan probe were designed to amplify the 81bp region on the cyt b gene. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct value obtained for Molva molva DNA (19.45±0.65) and the average Ct for non-target species DNA (38.3±2.8), even with closely related species such as Molva dypterygia (34.9±0.09). The proposed methodology has been validated with 31 commercial samples.

摘要

海鲜欺诈——通常涉及用一种物种替代另一种——由于在全球范围内普遍存在,已引起广泛关注。虽然DNA分析有助于打击这类欺诈行为,但目前使用的一些方法耗时且需要精密设备或训练有素的人员。这项工作描述了一种用于检测海产品中长鳍鳕(Molva molva)的新型实时荧光定量PCR TaqMan检测方法的开发。为此,设计了特异性引物和小沟结合(MGB)TaqMan探针,以扩增细胞色素b基因上的81bp区域。效率、特异性和交叉反应性分析表明,长鳍鳕(Molva molva)DNA获得的平均Ct值(19.45±0.65)与非目标物种DNA的平均Ct值(38.3±)之间存在统计学上的显著差异。2.8)之间存在统计学上的显著差异,即使是与长鳍鳕(Molva dypterygia)等近缘物种(34.9±0.09)也是如此。所提出的方法已在31个商业样品上得到验证。

(注:原文中“38.3±”后面似乎缺失数字,译文按原文翻译。)

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