Herein, a novel single-enzyme-assisted
dual recycle amplification
strategy based on T7 exonuclease (T7 Exo) and a strand-displacement
reaction (SDR) was designed to fabricate a photoelectrochemical (PEC)
biosensor for sensitive microRNA-141 (miRNA-141) detection with the
use of laminar bismuth tungstate (Bi2WO6) as
photoactive material. Compared with a traditional enzyme-assisted
dual recycle amplification strategy, the presented method could effectively
refrain the enzyme interference reaction, reduce environmental sensitivity,
and save cost. Here, hairpin DNA1 (H1) decorated on magnetic beads
(MB) hybridized with target miRNA-141 to form an H1/miRNA-141 heteroduplex.
With the introduction of hairpin DNA2 (H2)-labeled SiO2 (H2-SiO2), SDR was triggered between H2-SiO2 and H1, thus miRNA-141 was displaced from the H1/miRNA-141 heteroduplex
and an H1/H2-SiO2 duplex was formed, realizing the reuse
of the target. In the presence of T7 Exo, the H1/H2-SiO2 duplex was digested with the release of output DNA-SiO2. To enhance the target conversion rate, H1-MB was intactly released
and cycled, which could initiate more T7 Exo digestion and free abundant
output DNA-SiO2. Through such a process, a tiny miRNA-141
could induce substantial output DNA-SiO2, effectively improving
the target amplification efficiency and detection sensitivity of a
PEC biosensor. Furthermore, Bi2WO6 was modified
on an electrode to provide a superior initial PEC signal due to its
excellent electronic transformation capacity. With the introduction
of output DNA-SiO2, the hairpin structure of H3 on the
electrode was opened, making SiO2 close to the electrode
surface, which significantly decreases the PEC signal. This work first
established the PEC biosensor featuring a single-enzyme-assisted dual
recycle amplification process for sensitive detection of biomarkers.