MicroRNAs
(miRNAs) are useful biomarkers for the diagnosis of a
variety of cancers. However, it is a major challenge to detect miRNAs,
considering their high sequence similarity, low concentration, and
small size nature. With the establishment of an efficient rolling
circle amplification (RCA) molecular network by target-driven polymerization/nicking
reactions, we present here an exponential amplification strategy for
detecting miRNA in a label-free way with ultrahigh sensitivity. The
target miRNA sequences can bind two ssDNA probes to form a junction
structure to initiate a dual polymerization/nicking cyclic reaction
for the production of many primers, which further trigger multiple
RCA reactions in a drastically amplified sequence replication and
extension mode for the yield of substantial dsDNAs with various sizes.
The SYBR Green I then binds these dsDNAs to induce significantly magnified
fluorescence emission for detecting the target miRNA sequences with
a detection limit down to 0.86 fM in the linear range between 1 fM
and 10 pM. Because of the involvement of the presynthesized circular
DNA template, the RCA efficiency is further improved, and such a method
can also be used for detecting miRNA in diluted human serum samples,
demonstrating its great potential and universality for detecting different
nucleic acid sequences for biochemical research and clinical diagnosis
applications.
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