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

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.