中文摘要：堆积性状转基因生物因其生产效率的提高和功能特性的改进而日益普及，同时堆积性状转基因生物的现场检测成为一个迫切需要解决的挑战。本文开发了基于事件特异性TM-LAMP和脱氧核酶侧流式生物传感器的级联系统来可靠地检测堆积性状转基因生物（DP305423x GTS 40-3-2）。事件特异性TM-LAMP系统重新设计了三个引物集。脱氧核酶侧流式生物传感器在显示扩增产物的同时没有出现交叉污染，且脱氧核酶增加可以有效提高灵敏度。堆积性状转基因大豆的检测限约为0.1% (w/w)，且在没有任何大型仪器的情况下，整个检测过程可以缩短到120分钟。研究结果表明，这种方法不仅可以用于DP305423x GTS 40-3-2大豆单内核的现场检测，还可以在高度加工的食品中筛选堆积性状转基因大豆和亲本转基因大豆。
外文摘要：Stacked genetically modified organisms (GMO) are becoming popular for their enhanced production efficiency and improved functional properties, and on-site detection of stacked GMO is an urgent challenge to be solved. In this study, we developed a cascade system combining event-specific tag-labeled multiplex LAMP with a DNAzyme-lateral flow biosensor for reliable detection of stacked events (DP305423x GTS 40-3-2). Three primer sets, both event-specific and soybean species-specific, were newly designed for the tag-labeled multiplex LAMP system. A trident-like lateral flow biosensor displayed amplified products simultaneously without cross contamination, and DNAzyme enhancement improved the sensitivity effectively. After optimization, the limit of detection was approximately 0.1% (w/w) for stacked GM soybean, which is sensitive enough to detect genetically modified content up to a threshold value established by several countries for regulatory compliance. The entire detection process could be shortened to 120 min without any large-scale instrumentation. This method may be useful for the in-field detection of DP305423x GTS 40-3-2 soybean on a single kernel basis and on -site screening tests of stacked GM soybean lines and individual parent GM soybean lines in highly processed foods.
外文关键词：Stacked genetically modified soybean; Event-specific TM-LAMP; DNAzyme-lateral flow biosensor
作者：Cheng, Nan; Shang, Ying; Xu, Yuancong; 等
作者单位：China Agr Univ
期刊名称：BIOSENSORS & BIOELECTRONICS
期刊影响因子：7.476
出版年份：2017
出版刊次：5
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