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采用Taqman探针PCR技术,建立食品中大豆抗草甘膦转基因成分的定量检测方法。通过设计特异性引物,扩增内源基因lectin和35S启动子,建立两种基因的拷贝数-CT标准曲线,根据标准曲线方程计算样品中的转基因成分。结果表明,lectin和35S启动子基因标准曲线线性关系好,R2值分别为0.9934和0.9954,该方法对已知转基因成分含量的样品检测回收率为98%～128%,检测低限为0.025 ng。本方法具有快速、灵敏、准确、特异性高、等优点,可用于食品中大豆转基因成分的定量检测。
Taqman probe PCR technology was used to establish a quantitative method for the determination of glyphosate-resistant genetically modified soybeans in food. By designing specific primers, amplifying the endogenous gene lectin and 35S promoter, the copy number-CT standard curve of the two genes was established, and the transgenic components in the sample were calculated according to the standard curve equation. The results showed that the standard curve of lectin and 35S promoter had a good linear relationship, with R2 values of 0.9934 and 0.9954, respectively. The detection limit of this method was 98% -128% with the detection limit of 0.025 ng. The method has the advantages of fast, sensitive, accurate, high specificity and the like and can be used for the quantitative detection of genetically modified soy components in food.