In this work, a two-dimensional
self-assembled magnetic nanoparticle–graphene
oxide (MNP-GO) nanocomposite is reported for the detection of DNA.
Single-stranded DNA (ssDNA) coils, generated through a rolling circle
amplification (RCA) reaction triggered by the hybridization of target
oligos and padlock probes, have a strong interaction with MNP-GO nanotags
through several mechanisms including π–π stacking,
hydrogen bonding, van der Waals, electrostatic, and hydrophobic interactions.
This interaction leads to a hydrodynamic size increase or aggregation
of MNP-GO nanotags, which can be detected by a simple optomagnetic
setup. Due to the high shape anisotropy, MNP-GO nanotags provide stronger
optomagnetic signal than individual MNPs. Moreover, the avoidance
of DNA probes (i.e., short ssDNA sequences as the biosensing receptor)
provides easier material preparation and lower measurement cost. From
real-time measurements of interactions between MNP-GO and RCA products
amplified from a highly conserved Escherichia coli 16S rDNA sequence, a limit of detection of 2 pM was achieved with
a total assay time of 90 min. Although the nonspecific binding force
between GO and ssDNA is much weaker than the specific base-pairing
force in a DNA duplex, the proposed method provides a detection limit
similar to DNA probe-based magnetic biosensors, which can be ascribed
to the abundant binding sites between GO and ssDNA. In addition, for
target concentrations higher than 100 pM, the MNP-GO nanotags can
be applied for a qualitative naked eye detection strategy based on
nanotag–ssDNA flocculation.