Zettomole electrochemical HIV DNA detection using 2D DNA-Au nanowire structure, hemin/G-quadruplex and polymerase chain reaction multi-signal synergistic amplification

Highlights: • Two-dimensional DNA-Au nanowire was introduced to amplify the signal. • Novel PCR strategy effectively avoided the denaturation of PCR product. • Zettomole electrochemical detection of HIV DNA was realized (1.3 aM). • High sequence specificity obtained for HIV DNA detection. Multi-signal synergistically amplified electrochemical sensing of HIV DNA was proposed based on two-dimensional (2D) DNA-Au nanowire structure coupled with hemin/G-quadruplex and polymerase chain reaction (PCR). In the design, by using target HIV DNA as the template, PCR generated numbers of double-stranded DNA (dsDNA) with free single-stranded DNA (ssDNA) tails on one side and free G-quadruplex sequences on the other side. Then, the ssDNA tails of the PCR products were hybridized with the capture probe (CP) to introduce the hemin/G-quadruplex to the electrode surface as a redox-active reporter and to amplify the electrochemical signal as mimic peroxidase catalysis in the presence of H₂O₂. Meanwhile, (+)AuNPs were electrostatically adsorbed onto dsDNA surface for the formation of 2D DNA-Au nanowire structure, amplifying the electrochemical signal further as another mimic peroxidase and electric conductor together. By effectively combining these signal amplification processes, ultrasensitive HIV DNA detection was achieved with a detection limit of 1.3 aM, indicating that it has potential application in clinical diagnosis.