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

• Published in: Biosensors & bioelectronics 2017-11, Vol.97, p.26-33
• Main Authors: Zhang, Jian, Zhao, Qian, Wu, Yudong, Zhang, Bo, Peng, Weipan, Piao, Jiafang, Zhou, Yurui, Gao, Weichen, Gong, Xiaoqun, Chang, Jin
• Format: Article
• Language: English
• Subjects: Biosensing Techniques - methods; DNA - analysis; DNA - blood; DNA - genetics; Fluorescent Dyes - chemistry; Gold - chemistry; Gold-nanoparticles; Humans; Metal Nanoparticles - chemistry; Microbeads-based microchip; MicroRNAs - analysis; MicroRNAs - blood; Molecular beacons; Nanoparticle surface energy transfer; Nucleic Acid Hybridization - methods; Nucleic acids detection; Oligonucleotide Probes - chemistry; Polystyrenes - chemistry; Spectrometry, Fluorescence - methods; Thymidine Kinase - genetics
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2017.05.039

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 10min 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. •A novel gene detection microplatform for one-step detecting TK1-DNA and microRNA-21 was constructed.•The microplatform used NSET microbeads as nucleic acid probes.•The detection platform could detect the nucleic acid with high throughput, high sensitivity and high specificity.