MDA技术在转基因检测中的研究与应用

Development and Application of a Novel Genetically Modified Crops Detection Method Based on Multiple Displacement Amplification

PCR技术是转基因植物及其产品检测的主要方法,但是因为其对模板纯度和质量有较高要求,对于低质量模板难以获得稳定的检测结果。本研究利用MDA技术可将低至10 copies的微量DNA样品扩增至可用PCR方法稳定检出;进一步利用MDA技术对转基因玉米有证标准物质Bt11粉末的单颗粒痕量样品释出DNA进行扩增,结合PCR扩增检测,玉米粉末颗粒中内源参照基因的检出率达到70%以上,转基因特异性序列检出与内源参照基因检出的比值,与标准物质标称值基本相符,为建立基于MDA技术转基因植物及其产品粉末颗粒等痕量样品检测方法奠定了基础,同时也为加工产品中复合性状转基因作物的检测提供了潜在的解决方案。

PCR technology is the major genetically modified(GM) crops detection method, but the purity and quality of the template is the major limitation, furthermore, it is not suitable for the detection of the sample that has low purity and quality. In this paper, we develop a novel detection method based on multiple displacement amplification.The limit of detection reaches to 10 copies. More over, the method have also detected certified reference material of GM maize Bt11 by single particle derived from maize powder. The result showed the detection rate of endogenous reference gene is up to 70%. The ratio of the detected event specific sequence to endogenous reference gene was in agreement with the certified values. The results provide a novel method for detection of traceable GMsamples,meanwhile, it also provides a potential solution for the detection of stacked GM crops.