College of Chemistry and Material Science, Sichuan Normal University, Chengdu, Sichuan 610068, China.
J Mater Chem B. 2019 Aug 7;7(31):4778-4783. doi: 10.1039/c9tb00753a.
Due to their simplicity of design and operation, homogeneous bioassays have been of great interest to researchers. Herein, a label-free and free separation fluorescence sensing platform was constructed for the determination of nucleic acid and prostate specific antigen (PSA) using CdTe QDs as the signal molecule. In our previous work, we surprisingly found that the CdTe QDs can selectively distinguish Ag+ and the C-Ag+-C complex, which was the basis of the sensor. On the basis of the selective cation exchange reaction (CER), combined with the signal amplification of the strand displacement reaction (SDR), this work was first applied for the sensitive analysis of DNA. There are two types of hairpin structures in this sensing system, including the recognition probe (HP) and Ag+, which formed the C-Ag+-C structure, and the hairpin structure formed by the helper DNA itself. In this work, target DNA can trigger the SDR that generates lots of HP-helper double-stranded DNA (dsDNA) and recycles the target DNA while releasing a large amount of Ag+, thus quenching the fluorescence signal of CdTe QDs to achieve the highly sensitive detection of DNA. In order to verify the versatility of this system using DNA as a bridge and aptamers as recognition probes, we extended the system to the detection of PSA. After examining its experimental performance, it was determined that this method displayed good analytical capability for DNA in the range of 10-13-10-10 M and PSA in the range of 10-13-10-10 g mL-1 with low 25 fM and 30 fg mL-1 limits of detection (LODs), respectively; high selectivity for both the target sequence and protein was shown. In addition, this platform was successfully used for the analysis of PSA in serum samples.
由于其设计和操作简单,均相生物测定法引起了研究人员的极大兴趣。在此,我们构建了一种无标记且无需分离的荧光传感平台,用于使用 CdTe QD 作为信号分子测定核酸和前列腺特异性抗原 (PSA)。在我们之前的工作中,令人惊讶的是,我们发现 CdTe QD 可以选择性地区分 Ag+和 C-Ag+-C 配合物,这是传感器的基础。在此基础上,我们结合链置换反应 (SDR) 的信号放大,首次将选择性阳离子交换反应 (CER) 应用于 DNA 的灵敏分析。在这个传感系统中有两种发夹结构,包括识别探针 (HP) 和 Ag+,它们形成 C-Ag+-C 结构,以及由辅助 DNA 自身形成的发夹结构。在这项工作中,靶 DNA 可以触发 SDR,生成大量的 HP-helper 双链 DNA (dsDNA),同时回收靶 DNA 并释放大量 Ag+,从而猝灭 CdTe QD 的荧光信号,实现 DNA 的高灵敏检测。为了验证该系统以 DNA 为桥接物和适体作为识别探针的通用性,我们将该系统扩展到 PSA 的检测中。在检查了其实验性能后,确定该方法对 10-13-10-10 M 范围内的 DNA 和 10-13-10-10 g mL-1 范围内的 PSA 具有良好的分析能力,检测限分别为低至 25 fM 和 30 fg mL-1;对目标序列和蛋白质均表现出高选择性。此外,该平台成功地用于血清样品中 PSA 的分析。