3 research outputs found
Enzyme-Free Nucleic Acid Amplification Assay Using a Cellphone-Based Well Plate Fluorescence Reader
Nucleic acids, DNA
and RNA, provide important fingerprint information
for various pathogens and have significant diagnostic value; however,
improved approaches are urgently needed to enable rapid detection
of nucleic acids in simple point-of-care formats with high sensitivity
and specificity. Here, we present a system that utilizes a series
of toehold-triggered hybridization/displacement reactions that are
designed to convert a given amount of RNA molecules (i.e., the analyte)
into an amplified amount of signaling molecules without any washing
steps or thermocycling. Fluorescent probes for signal generation were
designed to consume products of the catalytic reaction in order to
push the equilibrium and enhance the assay fold amplification for
improved sensitivity and reaction speed. The system of toehold-assisted
reactions is also modeled to better understand its performance and
capabilities, and we empirically demonstrate the success of this approach
with two analytes of diagnostic importance, i.e., influenza viral
RNA and a micro RNA (miR-31). We also show that the amplified signal
permits using a compact and cost-effective smartphone-based fluorescence
reader, an important requirement toward a nucleic-acid-based point-of-care
diagnostic system