肉制品中猪源性成分相对定量检测方法研究

目的建立一种肉制品中猪源性成分的相对定量检测方法。方法针对猪的ER-beta基因的mRNA序列设计了特异性引物与探针序列,并对引物和探针的特异性进行了验证。构建重组质粒,利用质粒拷贝数的log值和Ct值构建标准曲线。通过标准曲线计算各个样本拷贝数,将待测样本和标准种源样本的拷贝数进行比较,确定检测样本的猪源性含量。结果实际掺加量分别为20%、40%、50%和60%的混合样品经该方法检测所得掺假量均值为猪体系24.8%、53.1%、53.1%、61%,检测结果与其实际含量基本一致。结论该方法基本能实现肉制品中猪源性成分的相对定量检测。

Study on relative quantitative detection method of porcine-derived components in meat products

Objective To establish a relative quantitative method for the detection of porcine-derived components in meat products. Methods Specific primers and probe sequences were designed for porcine ER-beta-based mRNA gene sequence, and the specificity of the primers and probes was verified. A recombinant plasmid was constructed, and a standard curve was constructed by the log value and Ct value of the plasmid copy number. The copy number of each sample was calculated by the standard curve, the copy number of the sample was compared to be tested with the standard provenance sample, and the porcine-derived content of the test sample was determined. Results The average adulteration of the mixed samples with actual addition amounts of 20%, 40%, 50% and 60% by the method were 24.8%, 53.1%, 53.1%, 61% of the pig system. The test results were basically the same as their actual content. Conclusion This method basically can realize the relative quantitative detection of the porcine-derived components in the meat product.