Departamento de Química Física y Analítica, Universidad de Oviedo, 33006 Oviedo, Spain.
Departamento de Química, Centro de Ciências da Natureza, Universidade Federal do Piauí, Teresina 64049-550PI, Brazil.
Sensors (Basel). 2017 Apr 18;17(4):881. doi: 10.3390/s17040881.
The design of screening methods for the detection of genetically modified organisms (GMOs) in food would improve the efficiency in their control. We report here a PCR amplification method combined with a sequence-specific electrochemical genosensor for the quantification of a DNA sequence characteristic of the 35S promoter derived from the cauliflower mosaic virus (CaMV). Specifically, we employ a genosensor constructed by chemisorption of a thiolated capture probe and -aminothiophenol gold surfaces to entrap on the sensing layer the unpurified PCR amplicons, together with a signaling probe labeled with fluorescein. The proposed test allows for the determination of a transgene copy number in both hemizygous (maize MON810 trait) and homozygous (soybean GTS40-3-2) transformed plants, and exhibits a limit of quantification of at least 0.25% for both kinds of GMO lines.
用于检测食品中转基因生物体（GMOs）的筛选方法的设计将提高其控制效率。我们在此报告了一种聚合酶链反应（PCR）扩增方法，结合序列特异性电化学基因传感器，用于定量分析源自花椰菜花叶病毒（CaMV）的 35S 启动子的 DNA 序列特征。具体来说，我们采用一种基因传感器，通过将硫醇化捕获探针和 -氨基硫酚金表面化学吸附，在传感层中捕获未纯化的 PCR 扩增子，并与用荧光素标记的信号探针结合。该检测方法允许对半合子（玉米 MON810 性状）和纯合子（大豆 GTS40-3-2）转化植物中的转基因拷贝数进行测定，对于两种类型的 GMO 品系，其定量限至少为 0.25%。
