WEN Hong-tao,YANG Yang,DING Yi-jia,et al.Establishment of qPCR and 3D-dPCR Detection Method of Genetically Modified Soybean MON87701[J].Soybean Science,2020,39(02):212-219.[doi:10.11861/j.issn.1000-9841.2020.02.0212]
转基因大豆MON87701品系qPCR和3D-dPCR检测方法的建立
- Title:
- Establishment of qPCR and 3D-dPCR Detection Method of Genetically Modified Soybean MON87701
- Keywords:
- Genetically modified soybean; MON87701; Quantitative detection; qPCR; 3D-dPCR
- 文献标志码:
- A
- 摘要:
- 转基因大豆MON87701是由孟山都公司研发的商业化抗虫大豆品系，目前已在17个国家广泛应用。为建立适用于MON87701大豆的高效检测方法，本研究基于实时荧光定量PCR(real-time fluorescence quantitive PCR, qPCR)和芯片式数字PCR（3D digital PCR, 3D-dPCR）平台，对44种不同转基因材料进行测试，建立双重定量检测方法。研究结果显示：只在转基因大豆MON87701样品中获得了阳性结果，证实该方法中引物和探针组合具有较高的特异性。进一步对2%、0.9%、0.09%和0.02%不同含量的MON87701转基因大豆进行定量测试，建立的qPCR和3D-dPCR两种方法的定量限均达到0.09%，检出限均达到0.02%，两者并无明显差别，但由于3D-dPCR和qPCR相比不需要标准物质制作标准曲线，并且对DNA提取质量要求更低，因此使用起来更加便捷高效，为转基因检测提供了新的方法和思路。
- Abstract:
- Genetically modified (GM) soybean event MON87701 is a commercial insect resistant soybean developed by Monsanto company, which has been widely used in 17 countries. In order to established efficient detection method of MON87701, this study established duplex quantitative detection method based on real-time fluorescence quantitive PCR (qPCR) and 3D digital PCR (3D-dPCR) platforms and tested 44 different transgenic materials. The result showed: The positive results were only obtained in the samples of MON87701, which confirmed that the primer and probe combination in the method had high specificity. The LOQs of the both methods were 0.09% and the LODs were 0.02%. There was no significant difference between the two methods. However, 3D-dPCR method does not need reference materials and the requirement on DNA quality is lower, it is more convenient and efficient providing new methods and ideas for the GMO detection.
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备注/Memo
收稿日期：2019-06-04