The construction of a novel nucleic acids detection microplatform based on the NSET for one-step detecting TK1-DNA and microRNA-21

分子信标 核酸 核酸定量 胶体金 DNA 检出限 微珠(研究) 荧光 纳米颗粒 化学 纳米技术 组合化学 材料科学 色谱法 寡核苷酸 生物化学 量子力学 物理
作者
Jian Zhang,Qian Zhao,Yudong Wu,Bo Zhang,Weipan Peng,Jiafang Piao,Yurui Zhou,Weichen Gao,Xiaoqun Gong,Jin Chang
出处
期刊:Biosensors and Bioelectronics [Elsevier]
卷期号:97: 26-33 被引量:12
标识
DOI:10.1016/j.bios.2017.05.039
摘要

Microbeads-based microchip technology has become the potential for a new generation of nucleic acids detection in a high-throughput and sensitive manner. However the specificity and operational complexity limit the microchip applied in nucleic acids detection. Herein, in this work, we designed a kind of gold-nanoparticles coated polystyrene microbeads as microplatform conjugating with the molecular beacons as probes. Due to the nanoparticle surface energy transfer of gold-nanoparticles, the fluorescence of dye on one end of molecular beacons was effectively quenched. When the target nucleic acids existed, the fluorescence of dye was quickly “turn-on” with high sensitivity. Due to the nanoparticle surface energy transfer effect of gold-nanoparticles, the designed platform performed better sensitivity than traditional microbead-based detection methods and realized quickly detection within 10 min without purification steps. In addition, compared with the linear chain probes, the molecular beacons probes enabled higher specificity and wash-free operation. Through different dyes encoded, TK1-DNA and microRNA-21 were simultaneously detected in one step and finally quantified by flow cytometry. The proposed detection method was also capable of monitoring TK1-DNA and microRNA-21 levels in human serum. Our study provides the potential multidetection of DNA and RNA.
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