Taqman实时定量PCR技术检测大豆转基因成分

TaqMan real-time quantitative PCR system to detect soy prosessed foods containing genetically modified organisms

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。

To establish quantitative method to detect the roundup eady soybean components in food by using TaqMan real-time quantitative PCR technology. The endogenous lectin and 35S promoter genes were amplified through the design of specific primers for them. The transgenic ratio was then calculated according to their standard copies-CT linear grapHs. The results showed that the R2 values for lectin and 35S promoter genes are 0.9934 and 0.9954 respectively. The recovery rate of the method is between 98%-128% and the detecting limit is 0.025 ng. The method is suitable for quantifying the Roundup Ready soyben components in food with fast, sensitive, accurate and high specificity.