Ultrasensitive Detection of DNA via SI-eRAFT and in Situ Metalization Dual-Signal Amplification

In this work, we report a new amplification strategy based on electrochemically mediated reversible addition fragmentation chain transfer (eRAFT) and in situ metalization for electrochemical detection of DNA. First, peptide nucleic acid (PNA) probes were immobilized on the surface of the gold electrode, and when they hybridized with the target DNA, the chain transfer agent (CTA), 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPAD), of RAFT was connected to the PNA/DNA heteroduplex formed by the coordination bonding of Zr4+. Then glycosyloxyethyl methacrylates (GEMA) were assembled on the surface of the electrode by electrochemically mediated surface-initiated reversible addition fragmentation chain transfer (SI-eRAFT) to form a polymer-containing sugar glucose. Next, the o-hydroxyl groups on the polysaccharide molecular skeleton were oxidized to aldehyde groups by sodium periodate (NaIO4). The aldehyde groups generated then reduce silver ions to silver particles deposited on the electrode surface in situ, and this system was then subjected to differential pulse voltammetry (DPV). Under optimal conditions, the intensity of the stripping current and the logarithm of the target DNA (tDNA) concentration has a good linear relationship in the range of 10 aM to 1 pM (R-2 = 0.996), and the detection limit can go down to 5.4 aM (S/N = 3). Moreover, the method is suitable for single-nucleotide polymorphism (SNP) analysis and has strong anti interference ability for the analysis of target ssDNA in serum samples.