title: Exponential strand-displacement amplification for detection of microRNAs.
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 Nt.AlwI, 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.
authors.affiliation: State Key Laboratory Base of Eco-chemical Engineering, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology , Qingdao, Shandong 266042, P. R. China.
authors.forename: Chao
authors.initials: C
authors.lastname: Shi
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.forename: Cuiping
authors.initials: C
authors.lastname: Ma
authors.forename: Wenwan
authors.initials: W
authors.lastname: Zhong
mesh_terms: D006801:Humans; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D035683:MicroRNAs