Exponential Strand-Displacement Amplification for
Detection of MicroRNAs
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Abstract
MicroRNAs
(miRNAs) are promising targets for disease diagnosis. However, miRNA
detection requires rapid, sensitive, and selective detection to be
effective as a diagnostic tool. Herein, a miRNA-initiated exponential
strand-displacement amplification (SDA) assay was reported. With the
Klenow fragment, nicking enzyme <i>Nt.AlwI</i>, and two
primers, the miRNA target can trigger two cycles of nicking, polymerization,
and displacement reactions. These reaction cycles amplified the target
miRNA exponentially and generated dsDNAs detectable with SYBR Green
I in real-time PCR. As low as 16 zmol of the target miRNA was detected
by this one-pot assay within 90 min, and the dynamic range spanned
over 9 orders of magnitude. Negligible impact from the complex biological
matrix was observed on the amplification reaction, indicating the
assay’s capability to directly detect miRNAs in biofluids