An isothermal, non-enzymatic, and dual-amplified fluorescent sensor for highly sensitive DNA detection

Abstract Sensitive DNA assays are of importance in life science and biomedical engineering, but they heavily dependent on thermal cycling programs or enzyme-assisted schemes, which require the utilization bulky devices costly reagents. To circumvent such requirements, we developed an isothermal enzyme-free sensing method with dual-stage signal amplification ability based coupling use catalytic hairpin assembly (CHA) Mg 2+ -dependent deoxyribozyme (DNAzyme). In this study, system involves a set probes for CHA (ensuring first stage amplification) as well ribonucleobase-modified molecular beacons that serve activatable substrates DNAzymes (warranting second amplification). An experimentally determined detection limit about 0.5 pM is achieved good linear range from to 10 pM. The results spiked fetal bovine serum samples further confirm reliability practical applications. non-thermal cycling, enzyme-free, dual-amplified features make it powerful tool effective nucleic acid assay variety