Department of Mechanical Engineering, National Chung-Hsing University, Taichung 40227, Taiwan.
Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung 40227, Taiwan.
Biosensors (Basel). 2022 Mar 30;12(4):207. doi: 10.3390/bios12040207.
In this study, we proposed a genosensor that can qualitatively and quantitatively detect genetically modified soybeans using a simple electrode with evenly distributed single layer gold nanoparticles. The DNA sensing electrode is made by sputtering a gold film on the substrate, and then sequentially depositing 1,6-hexanedithiol and gold nanoparticles with sulfur groups on the substrate. Then, the complementary to the CaMV 35S promoter (P35S) was used as the capture probe. The target DNA directly extracted from the genetically modified soybeans rather than the synthesized DNA segments was used to construct the detection standard curve. The experimental results showed that our genosensor could directly detect genetically modified genes extracted from soybeans. We obtained two percentage calibration curves. The calibration curve corresponding to the lower percentage range (1-6%) exhibits a sensitivity of 2.36 Ω/% with R = 0.9983, while the calibration curve corresponding to the higher percentage range (6-40%) possesses a sensitivity of 0.1 Ω/% with R = 0.9928. The limit of detection would be 1%. The recovery rates for the 4% and 5.7% GMS DNA were measured to be 104.1% and 102.49% with RSD at 6.24% and 2.54%. The gold nanoparticle sensing electrode developed in this research is suitable for qualitative and quantitative detection of genetically modified soybeans and can be further applied to the detection of other genetically modified crops in the future.
在这项研究中，我们提出了一种基因传感器，它可以使用带有均匀分布单层金纳米粒子的简单电极定性和定量检测转基因大豆。DNA 传感电极是通过在基底上溅射金膜，然后在基底上依次沉积带有硫基团的 1,6-己二硫醇和金纳米粒子制成的。然后，使用与 CaMV 35S 启动子（P35S）互补的序列作为捕获探针。该检测标准曲线使用的是直接从转基因大豆中提取的目标 DNA，而不是合成的 DNA 片段。实验结果表明，我们的基因传感器可以直接检测从大豆中提取的转基因基因。我们得到了两个百分比校准曲线。对应于较低百分比范围（1-6%）的校准曲线具有 2.36 Ω/%的灵敏度，R = 0.9983，而对应于较高百分比范围（6-40%）的校准曲线具有 0.1 Ω/%的灵敏度，R = 0.9928。检测限为 1%。测量 4%和 5.7% GMS DNA 的回收率分别为 104.1%和 102.49%，RSD 为 6.24%和 2.54%。本研究开发的金纳米粒子传感电极适用于转基因大豆的定性和定量检测，并可进一步应用于未来其他转基因作物的检测。
