JiangXi University of Chinese Medicine, Nan Chang, JiangXi, 330004, China.
JiangXi University of Chinese Medicine, Nan Chang, JiangXi, 330004, China.
Biosens Bioelectron. 2023 Sep 1;235:115385. doi: 10.1016/j.bios.2023.115385. Epub 2023 May 9.
Herein, an fluorescence (FL)-electrochemiluminescence (ECL) dual-mode biosensor is constructed based on the dual-signal "turn-on" strategy of functionalized metal-organic frameworks nanosheets (RuMOFNSs)-tetraferrocene for K-ras gene detection, and the mechanism of bursting through front-line orbital theory is explained for the first time. Amino-functionalized tetraferrocene-labeled probe DNA molecules are linked to RuMOFNSs by covalent amide bonds, acting as FL and ECL intensity switches. The target DNA, complementary to the probe DNA, triggers cyclic amplification of the target by nucleic acid exonuclease III (Exo III), repelling tetraferrocene reporter groups away from RuMOFNSs and inhibiting the electron transfer process and photoinduced electron transfer (PET) effect. These phenomena induce a double turn-on of FL and ECL signals with a high signal-to-noise ratio. The developed FL-ECL dual-mode sensing platform provides sensitive detection of the K-ras gene with detection limits of 0.01 fM (the detection range is 1 fM to 1 nM) and 0.003 fM (the detection range is 0.01 fM to 10 pM), respectively. In addition, the proposed dual-mode sensor can be easily extended to detect other disease-related biomarkers by changing the specific target and probe base sequences, depicting potential applications in bioanalysis and early disease diagnosis.
本文基于功能化金属有机骨架纳米片(RuMOFNSs)-四聚体的双信号“开启”策略,构建了一种荧光(FL)-电化学发光(ECL)双模生物传感器,用于 K-ras 基因检测,并首次通过前线轨道理论解释了爆发机制。经氨基功能化的四聚体标记探针 DNA 分子通过共价酰胺键与 RuMOFNSs 连接,充当 FL 和 ECL 强度开关。与探针 DNA 互补的靶 DNA 通过核酸外切酶 III(Exo III)触发靶分子的循环扩增,将四聚体报告基团从 RuMOFNSs 上排斥开,并抑制电子转移过程和光诱导电子转移(PET)效应。这些现象导致 FL 和 ECL 信号的双重开启,具有高信噪比。所开发的 FL-ECL 双模传感平台对 K-ras 基因进行了灵敏检测,检测限分别为 0.01 fM(检测范围为 1 fM 至 1 nM)和 0.003 fM(检测范围为 0.01 fM 至 10 pM)。此外,通过改变特定的靶标和探针碱基序列,该双模传感器可以很容易地扩展到检测其他与疾病相关的生物标志物,描绘了在生物分析和早期疾病诊断中的潜在应用。