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随着全球商品化生产的转基因产品日益增多,转基因品系检测已逐渐成为国内及国际上转基因成分检测的发展趋势。为实施我国转基因农产品的进出境、环境释放及标签标识等相关法规,实现进口转基因大豆(Glycine max)的有效管理,本研究针对DAS-81419-2、DP356043-5、A2704-12、FG72、BPSCV127-9和MON89788等6种常见转基因大豆的旁侧序列,分别设计了环介导等温扩增(loop-mediated isothermal amplification,LAMP)引物。通过引物筛选及方法的特异性、灵敏度及检出限验证,建立了转基因大豆DAS-81419-2、DP-356043、A2704-12、FG72、BPS-CV127-9和MON89788的LAMP检测方法。结果表明,所建立的方法能特异、稳定地检测转基因大豆DAS-81419-2、DP356043-5、A2704-12、FG72、BPSCV127-9和MON89788,检测灵敏度可达到0.1%(W/W),通过在反应终产物中加入SYBR GreenⅠ染料,用肉眼即可直接判断检测结果,检测时间在30 min以内。本方法快速准确,且不需要复杂昂贵的仪器设备,适用于检测机构及相关执法部门对转基因产品的快速检测。
With the increasing global commercial production of genetically modified products, testing of transgenic lines has gradually become the development trend of detection of genetically modified organisms both at home and abroad. In order to realize the effective management of imported Glycine max for implementing the import and export, environmental release and labeling of genetically modified agricultural products in China, this study aimed at the effective management of imported Glycine max. -9 and MON89788 were used to design loop-mediated isothermal amplification (LAMP) primers. The method of LAMP detection of transgenic soybean DAS-81419-2, DP-356043, A2704-12, FG72, BPS-CV127-9 and MON89788 was established by primer screening and the specificity, sensitivity and detection limit detection. The results showed that the established method could detect the sensitivity and specificity of transgenic soybeans DAS-81419-2, DP356043-5, A2704-12, FG72, BPSCV127-9 and MON89788 with 0.1% (W / W) SYBR GreenⅠ dye was added to the final product of the reaction, which can be directly judged by the naked eye. The detection time is within 30 min. The method is rapid and accurate, does not require complicated and expensive equipment, and is suitable for rapid detection of genetically modified products by testing organizations and relevant law enforcement agencies.