A modified non-cross-linking gold-nanoparticles (Au-NPs) aggregation strategy has been developed for the label free colorimetric detection of DNAs/RNAs based on self-assembling target species in the presence of thiolated probes. Two complementary thiol- modified probes, each of which specifically binds at one half of the target introduced SH groups at both ends of dsDNA. Continuous disulfide bond formation at 3' and 5' terminals of targets leads to the self-assembly of dsDNAs into the sulfur- rich and flexible products with different lengths. These products have a high affinity for the surface of Au-NPs and efficiently protect the surface from salt induced aggregation. To evaluate the assay efficacy, a small part of the citrus tristeza virus (CTV) genome was targeted, leading to a detection limit of about 5 × 10-9 mol.L-1 over a linear ranged from 20 × 10-9 to 10 × 10-7 mol.L-1. This approach also exhibits good reproducibility and recovery levels in the presence of plant total RNA or human plasma total circulating RNA extracts. Self-assembled targets can be then sensitively distinguished from non-assembled or mismatched targets after gel electrophoresis. The disulfide reaction method and integrating self-assembled DNAs/RNAs targets with bare AuNPs as a sensitive indicator provide us a powerful and simple visual detection tool for a wide range of applications.

译文

:已开发出一种改进的非交联金纳米粒子(Au-NPs)聚集策略,用于在硫醇化探针存在下基于自组装靶物种的DNA / RNA的无标记比色检测。两个互补的巯基修饰探针,每个探针都与dsDNA两端的目标导入的SH基团的一半特异性结合。在靶标的3'和5'末端连续形成二硫键,导致dsDNA自组装成具有不同长度的富硫和柔性产物。这些产品对Au-NPs的表面具有很高的亲和力,并有效地保护了表面免于盐诱导的聚集。为了评估测定的功效,针对了小部分的柑桔类Tristeza病毒(CTV)基因组,在20 limit×10-9至10的线性范围内导致检出限约为5×10-9 mol.L-1。 ×10-7 mol.L-1。在存在植物总RNA或人血浆总循环RNA提取物的情况下,该方法还具有良好的重现性和回收率。凝胶电泳后,可以将自组装的靶标与未组装或错配的靶标区分开。二硫键反应方法以及将自组装的DNA / RNA靶标与裸金纳米颗粒结合在一起作为敏感指示剂,为我们提供了功能强大且简单的视觉检测工具,可用于广泛的应用。

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