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Identification of European Hake Species (Merluccius merluccius) Using Real-Time PCR

机译:使用实时荧光定量PCR鉴定欧洲无须鳕种(Merluccius merluccius)

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摘要

A rapid and precise method for identifying European hake (Merluccius merluccius) based on TaqMan technology is presented. The method can be applied to fresh, frozen, and processed fish products to detect the fraudulent or unintentional mislabeling of this species. Specific primers and a minor groove binding (MGB) TaqMan probe were designed for this purpose based on partial sequences of the mitochondrial DNA control region. Combinations of primers and probe concentrations that gave the lowest Ct value and the highest final fluorescence value were selected to carry out efficiency, specificity, and cross-reactivity assays. The method was successfully tested on 31 commercial hake samples. A Ct value of about 16 was obtained when Merluccius merluccius was present; however, the fluorescence signal was not detected most of the time (Ctvalue 40) or presented significantly higher Ct values (38.2 ± 0.96) for the nonhake species.
机译:提出了一种基于TaqMan技术的快速准确的欧洲无须鳕(Merluccius merluccius)识别方法。该方法可以应用于新鲜,冷冻和加工的鱼制品,以检测该种类的欺诈性或无意识的错误标签。为此,基于线粒体DNA控制区的部分序列,设计了特异性引物和小沟结合(MGB)TaqMan探针。选择给出最低Ct值和最高最终荧光值的引物和探针浓度的组合来进行效率,特异性和交叉反应性测定。该方法已在31种商业鳕鱼样品上成功测试。存在淡水默鲁氏菌时,Ct值约为16。然而,对于大多数非杂种而言,大部分时间都未检测到荧光信号(Ctvalue 40)或呈现出更高的Ct值(38.2±0.96)。

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