转基因抗虫玉米浙大瑞丰8实时荧光定量PCR检测方法
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农业农村部农业国家和行业标准制修订项目(农质标函〔2022〕66号); 科技创新2030重大项目(2022ZD0401909); 农业科研杰出人才培养计划


Real-time fluorescent quantitative PCR for detection of transgenic insect-resistant maize Zheda Ruifeng 8
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    摘要:

    【目的】 转基因抗虫玉米浙大瑞丰8是杭州瑞丰生物科技有限公司研发的新型抗虫玉米,于2021年获批农业转基因生物安全证书(生产应用)。本研究旨在为转基因抗虫玉米浙大瑞丰8设计特异性扩增引物和TaqMan探针,并对其转化体成分进行定量检测,为管理部门提供安全监管的依据。【方法】 通过体系优化、正确度及精密度检测、稳健性测试等系列研究,设计特异性引物和探针,对浙大瑞丰8转化体成分进行定量检测。【结果】 本研究研制的方法能够特异、定量检测出浙大瑞丰8转化体成分,对拷贝数比值在0.1% (20个拷贝数)以上的阳性样品可进行准确定量,对0.05% (10个拷贝数)的阳性样品可特异性检出。【结论】 本研究结果为管理部门对浙大瑞丰8的安全监管提供了有效的支撑,说明研制的方法可用于浙大瑞丰8的定量检测和安全监管。

    Abstract:

    【Aim】 Genetically modified (GM) insect-resistant maize, Zheda Ruifeng 8, is a new type of insect-resistant maize developed by Hangzhou Ruifeng Biotechnology Co. Ltd., and it was approved for an agricultural genetically modified organism safety certificate (for production and application) in 2021. This study aimed to design specific amplification primers and TaqMan probes for the transgenic insect-resistant maize Zheda Ruifeng 8 and to quantitatively detect its transgenic components, thereby providing a basis for safety supervision by regulatory authorities. 【Method】 Through a series of experiments, including system optimization, accuracy and precision testing, and robustness testing, specific primers and probes were designed to quantitatively detect the transgenic components of Zheda Ruifeng 8. 【Result】 The data showed that the method can specifically and quantitatively detect the transgenic components of Zheda Ruifeng 8. Positive samples with a copy number ratio of >0.1% (20 copies) were accurately quantified, and positive samples with a copy number ratio of 0.05% (ten copies) were specifically detected. 【Conclusion】 The findings of this study provide effective support for safety supervision by regulatory authorities, thereby indicating that the developed method can be used for the quantitative detection and safety supervision of Zheda Ruifeng 8.

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田锦,句荣辉,蒋红叶,陈子言,张华,李凌燕,孙宇,王颢潜,陈红,梁晋刚.转基因抗虫玉米浙大瑞丰8实时荧光定量PCR检测方法[J].生物安全学报中文版,2025,34(2):130-136

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  • 收稿日期:2024-02-05
  • 最后修改日期:2024-07-01
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  • 在线发布日期: 2025-05-16
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