二重荧光定量PCR检测转基因大豆DAS81419

Multiplex real-time PCR for the detection of genetically modified soybean DAS81419

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。

Specific primers and probes based on the 5＇ flanking sequence of exogenous fragments of genetically modified （GM） soybean DAS81419 and endogenous Lectin gene of soybean were designed. A multiplex real-time PCR for the detection of GM soybean DAS81419 and endogenous Lectin gene of soybean simultaneously has been developed. The specificity of this method has been tested by 15 GM soybean,3 GM maize,1 GM rape,1 GM rice and 1 non-GM soybean. DAS81419 was used to prepare 6 content gradients to test the sensitivity. The results show that this multiplex real-time PCR method was of high specificity for the detection of GM soybean DAS81419. The sensitivity of this method was 0.01%. In conclusion,this multiplex real-time PCR method is high specific,sensitive and reliable,and will be an effective tool for the detection of GM DAS81419 in the exit and entry inspection laboratory.