转基因玉米MON87411实时荧光PCR定性检测方法的建立及其标准化

Establishment and Standardization of Real-time PCR Method for Qualitative Detection of Genetically Modified Maize MON87411

转基因玉米MON87411是孟山都远东有限公司研发的抗虫耐除草剂玉米转化体,该转化体已获得进口加工原料的农业转基因生物安全证书。以转基因玉米MON87411品系特异性序列为靶标设计引物和探针,经特异性测试、体系优化、灵敏度测试及检出限测试,建立了转基因玉米MON87411的实时荧光PCR定性检测方法。结果表明,该方法能检测出转基因玉米MON87411的转化体成分,检出限(limit of detection,LOD)可达0.05%,具有稳定性好、特异性强、灵敏度高的特点。国内8家转基因生物安全检测机构对本方法进行了特异性测试、检出限测试和再现性测试,循环验证报告显示该方法符合国家标准方法的各项要求,可在检测行业推广应用。研究建立的方法可为我国对转基因玉米MON87411品系的安全监管提供有效的技术支撑。

Genetically modified(GM)maize MON87411 is an event developed by Monsanto,and it has obtained the biological safety certificate imported as raw materials for processing.In this paper,the primers and probes were designed with the specific sequences as targets.Through specificity test,system optimization,sensitivity test and limit of detection(LOD)test,a real-time PCR method for the qualitative detection of GM maize MON87411 was established.The results showed that the method could de‐tect the components of GM maize MON87411,and had the characteristics of good stability,strong specificity and high sensitivity and the LOD could reach 0.05%.The specific test,LOD test and reproducibility test of the method were carried out by 8 domes‐tic GMO safety testing institutions.The cyclic verification report showed that the method met the requirements of the national standard method and could be promoted and applied in the testing industry.The establishment of this method could provide effec‐tive technical support for the safety supervision of GM maize MON87411 strain in China.