The
unique discriminating ability of hydroxyapatite (HAP) to single-
and double-stranded nucleic acid implies its potential as a biosensing
material for nucleic acid analysis. Herein, a novel immobilization-
and label-free electrochemical strategy has been developed for miRNA
let-7a detection based on the specific adsorption of magnetic Fe3O4@hydroxyapatite (Fe3O4@HAP)
to a duplex structure. Typically, the highly efficient homogeneous
reaction between probe DNA (pDNA) and target miRNA let-7a produces
the nucleic acid duplex, which is specifically adsorbed on the surface
of Fe3O4@HAP. Thereafter, the electroactive
intercalator Nile blue (NB) is introduced as the label-free signal
molecule and inserts between the stacked base pairs of the pDNA–miRNA
duplex. Thus, the Fe3O4@HAP bearing pDNA–miRNA
and numerous intercalated NBs are adsorbed by the magnetic glassy
carbon electrode, giving rise to an intense sensing signal without
the need of nuclease-assisted cyclic amplification. With this advantage,
the sensing system shows a wide linear range from 1.0 fM to 100 nM
and a low detection limit of 0.051 fM for miRNA let-7a analysis. What
is more, the synergy of the base-pairing-driven hybridization, the
specific intercalation of NB to the duplex structure, and the selective
adsorption of Fe3O4@HAP contribute to a high
sensing specificity of the biosensing strategy. The sensing method
provides a state-of-the-art strategy for the sensitive and facile
bioanalysis and clinical diagnosis of miRNA.