Sensitive molecular diagnostics using volume-amplified magnetic nanobeads.

Abstract

In this letter, we demonstrate a new principle for diagnostics based on DNA sequence detection using single-stranded oligonucleotide tagged magnetic nanobeads. The target DNA is recognized and volume-amplified to large coils by circularization of linear padlock probes through probe hybridization and ligation, followed by rolling circle amplification (RCA). Upon hybridization of the nanobeads in the RCA coils, the complex magnetization spectrum of the beads changes dramatically, induced by the attached volume-amplified target molecules. We show that the magnetization spectrum of the nanobeads can be used for concentration determination of RCA coils down to the pM range, thus creating the opportunity for nonfluorescence-based cost-efficient high-sensitivity diagnostics tool. We also show that the bead incorporation in the coils is diffusion-controlled and consequently may be accelerated by incubating the sample at higher temperatures.