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本文针对大豆内源基因Lectin和转基因大豆DAS81419品系的5′端插入位点序列,设计特异性引物及探针,建立了同时检测转基因大豆DAS81419品系和大豆内源基因Lectin的二重荧光定量PCR方法,运用15种转基因大豆、3种转基因玉米、1种转基因油菜、1种转基因水稻和非转基因大豆对该方法进行了特异性评价,并分析了该方法的灵敏度和稳定性。结果显示,该方法能准确从20种转基因样品和1种非转基因样品中检出靶目标,检测结果与待检样品信息一致,表明本方法具有良好的特异性;灵敏度高达0.01%;并具有良好的重复性。该方法特异性强、灵敏度高、稳定性强,适用于各口岸实验室进行转基因大豆DAS81419的快速、准确的检测。
In this paper, specific primers and probes were designed based on the 5 ’end sequences of endogenous Lectin and DAS81419 in transgenic soybean, and a dual-fluorescence quantitative PCR method was established for the simultaneous detection of transgenic soybean DAS81419 and soybean endogenous gene Lectin The method was evaluated by using 15 transgenic soybean, 3 transgenic maize, 1 transgenic rape, 1 transgenic rice and non-genetically modified soybean. The sensitivity and stability of this method were also analyzed. The results show that the method can accurately detect target from 20 transgenic samples and 1 non-transgenic sample. The results are in good agreement with the information of samples to be tested, indicating that this method has good specificity; the sensitivity is up to 0.01%; and the method has good Repeatability. The method has the advantages of high specificity, high sensitivity and strong stability, and is applicable to the rapid and accurate detection of transgenic soy DAS81419 in various port laboratories.