随着我国转基因玉米和转基因大豆的商业化种植，为了保证我国生物育种产业的健康发展，迫切需要开发转基因玉米和大豆的现场快速检测技术。本研究针对转基因玉米和大豆现场快速检测技术的不足，提出了一种基于重组酶聚合酶扩增技术(recombinase polymerase amplification，RPA)的快速检测方法。选择具有代表性的转基因玉米(双抗12-5、DBN9936、MON810)和大豆(DBN9004、GTS40-3-2、SHZD32-1)为检测对象，根据其侧翼序列，筛选特异性引物和探针，建立了高特异性、灵敏度达20拷贝的RPA扩增体系。为了快速获取DNA，种子样品利用核酸释放剂获得可用于RPA扩增的模板，为了消除叶片中叶绿素并快速获取DNA，叶片样品利用本实验室研发的核酸快速一体化提取装置快速获取DNA，以消除叶绿素干扰。结合便携式金属浴启动RPA扩增，蓝光切胶仪用于可视化检测，构建了转基因玉米和大豆的现场快速检测方法。该方法检测时间约为20 min，与常规PCR检测结果一致，适用于现场快速检测，无需大型设备，具备较强的实用性和适应性。该方法为转基因作物现场检测提供了技术支持，并为其他核酸快速检测技术的应用提供参考。

With the commercial planting of transgenic maize and transgenic soybeans in China, in order to ensure the healthy development of China’s biological breeding industry, it is urgent to develop on-site rapid detection techniques for transgenic maize and soybeans. In this study, a rapid detection method based on recombinase polymerase amplification (RPA) was proposed to overcome the shortcomings of on-site rapid detection of transgenic maize and soybean. Representative transgenic maize (Shuang Kang 12-5, DBN9936, MON810) and soybean (DBN9004, GTS40-3-2, SHZD32-1) were selected as test objects. According to their flanking sequence, specific primers and probes were designed and screened, and a RPA amplification system with high specificity and sensitivity of 20 copies for each transgenic event was established. In order to quickly obtain DNA, seed samples use nucleic acid release agents to obtain templates that can be used for RPA amplification. In order to eliminate the interference of chlorophyll in leaves on RPA fluorescence signal and quickly obtain DNA, leaf samples use the nucleic acid rapid integrated extraction device developed by our laboratory to quickly obtain DNA and eliminate the chlorophyll interference. Combined with a portable metal bath for initiating RPA amplification, a blue light gel cutter for visual detection, and a rapid on-site detection method for transgenic maize and soybean was constructed. The entire detection process required approximately 20 minutes, and the detection result was consistent with the result of conventional PCR. It is suitable for on-site rapid detection without large equipment, and has strong practicability and adaptability. The proposed approach provides technical support for on-site detection of transgenic crops and offers a reference for the application of other rapid nucleic acid detection technologies.