Catalytic hairpin assembly-assisted lateral flow assay for visual determination of microRNA-21 using gold nanoparticles

Abstract

The authors describe an improved lateral flow assay based on (a) the use of catalytic hairpin assembly (CHA), and (b) on signal amplification performed at the interface of gold nanoparticles (AuNPs). The combination of the amplification capability of the CHA reaction and the unique optical properties of AuNPs results in an assay that has a sensitivity that is improved by more than two orders of magnitude. MicroRNA-21 was employed as a model analyte to prove the concept. The presence of microRNA-21 triggers the self-assembly of two hairpin DNAs into double stranded DNA and exposing biotin molecules on the surface of AuNPs. Hence, the target becomes recycled and the signal is strongly amplified. The AuNPs carrying biotin are captured on the test line of the strip to display a red zone. This enables the visual recognition of microRNA without the need for any instrumentation. The fast quantitation of microRNA via the red band intensity is accomplished with the help of software, and the limit of detection is 0.89 pM. The enhanced lateral flow assay was employed to the determination of microRNA-21 in cell extracts and spiked serum samples. Graphical abstract A lateral flow assay for microRNA is described with a detection limit that is improved by two orders of magnitude. It is based on catalytic hairpin assembly (CHA) signal amplification performed at the interface of gold nanoparticles.

Keywords: Cancer cells; DNA nanotechnology; Hairpin probes; Human serum; Quantification; Signal amplification.