Surface plasmon resonance biosensor for enzyme-free amplified microRNA detection based on gold nanoparticles and DNA supersandwich

A novel enzyme-free amplified surface plasmon resonance (SPR) biosensor for microRNA (miRNA) detection was developed based on gold nanoparticles (AuNPs) coupled with DNA supersandwich. In the detection strategy, the DNA-linked AuNPs as the primary amplification element, not only hybridized with the capture DNA on the Au film to amplify SPR signal but also initiated the subsequent secondary amplification, i.e. DNA supersandwich formation of two report probes. In the presence of target, stem-loop structure of capture DNA on the Au film surface was unfolded, and DNA-linked AuNPs were bound to Au film by hybridization with terminus of capture DNA. Then, the carried assistant DNA on the AuNPs could trigger an alternative hybridization reaction of two report probes, resulting in the formation of DNA supersandwich. Due to the electronic coupling between localized plasmon of AuNPs and the surface plasmon wave associated with Au film, as well as the enhancement of the refractive index of the medium next to the metal film caused by DNA supersandwich structure, the shift of resonance angle was enhanced obviously. By employing the enzyme-free dual signal amplification strategies, as low as ca. 8 IM miRNA-21 could be detected. Moreover, this assay also showed high selectivity toward single-base mismatch, and demonstrated its applicability for the target detection in human serum. This work may provide great potential applications in future clinical analysis. (C) 2015 Elsevier B.V. All rights reserved.