Ternary Electrochemiluminescence Biosensor Based on DNA Walkers and AuPd Nanomaterials as a Coreaction Accelerator for the Detection of miRNA-141

In this study, a ternary electrochemiluminescence (ECL) sensing platform coupled with a multiple signal amplification strategy was proposed for ultrasensitive detection of miRNA-141. The initial signal amplification was achieved via three-dimensional reduced graphene oxide (3D-rGO)@Au nanoparticles (NPs) to form an excellent conductive layer. Then, AuPd NPs as a coreaction accelerator was introduced into the N-(4-aminobutyl)-N-(ethylisoluminol) (ABEI)-H₂O₂ system to facilitate the transformation from H₂O₂ to excess superoxide anion radicals (O₂^•-), which further amplified the ECL emission of ABEI, leading to a significant increase of the ECL signal. Meanwhile, in the presence of miRNA-141 and T7 Exonuclease (T7 Exo), the self-assembled DNA swing arm can be driven to walk autonomously. The DNA walker reaction could result in the release of numerous labeled luminophores, which could react to achieve an extremely weak ECL signal. Surprisingly, the established ECL sensor platform for the detection of miRNA-141 demonstrated excellent sensitivity with a low detection limit of 31.9 aM in the concentration range from 100 aM to 1 nM. Consequently, the designed strategy greatly improves the luminous efficiency of the ternary ECL system and provides a special approach for the detection of nucleic acids and biomarkers in clinical and biochemical analysis.