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采用环状等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对转基因玉米品系MON863中玉米基因组与外源基因结合处设计4条特异性引物,建立MON863的快速检测技术体系,并对时间、温度等反应条件及体系灵敏度、特异性等进行探索。结果表明:该检测体系最适反应温度为63℃,反应时间60 min,灵敏度是常规定性PCR的10倍。该检测方法具有高度的特异性和稳定性,操作方便、简单、省时。
Four specific primers were designed for the combination of maize genome and foreign gene in transgenic maize MON863 by using Loop-Mediated Isothermal Amplification (LAMP). The rapid detection technology system of MON863 was established. The effects of time, Temperature and other reaction conditions and system sensitivity, specificity, etc. to explore. The results showed that the optimal reaction temperature was 63 ℃ and the reaction time was 60 min. The sensitivity was 10 times of that of conventional qualitative PCR. The detection method has a high degree of specificity and stability, easy to operate, simple, time-saving.