High-Specificity Kilobase-Level Multiplex PCR Enabled by Graphene Oxide-Dendrimer-Encapsulated Gold Nanoparticles

Abstract

To address the challenges of nonspecific amplification in kilobase (kb)-level multiplex polymerase chain reaction (PCR), we developed an innovative nanomaterial—graphene oxide-dendrimer-encapsulated gold nanoparticles (GO-G3-Au). This material achieves high amplification specificity and enables high-efficiency kb-level multiplex PCR to be completed within approximately 70 min, while significantly improving amplification uniformity (ΔCt between multiplex amplicons < 4), even under conditions where conventional methods fail. Furthermore, experimental analysis and theoretical studies using zeta potential analysis, thermal conductivity measurements and finite element simulations supported that the enhancement in PCR performance arises from the synergistic effects of interfacial enrichment and high thermal conductivity. Collectively, these findings establish GO-G3-Au as an effective platform for enhancing the specificity, uniformity and efficiency of nucleic acid amplification, thereby advancing molecular diagnostics.