Nucleic acid signal amplification technologies and their cascade systems possess powerful ability to amplify signals. However, their application in biological computing, which necessitates a strict logical relationship, requires further investigation. To address this need, we developed a flexible sensing substrate consisting of carbon fiber cloth/graphene walls (CFC/GW), integrated with a 3D DNA tetrahedron (DNA-Te)-assisted bioplatform. This platform serves as the foundation for two types of Boolean logic gates: the “AND” and “OR” gate. The output system of the Cu2+ + DNAzyme logic gate module, activated by multiple miRNAs, was combined with the CFC/GW/Au-Te-based detection system to precisely identify small extracellular vesicle (sEV)-derived miRNAs in ultratrace quantities. Leveraging DNA Te-assisted DNAzyme molecular logic circuits, we designed higher-order Boolean logic operations to effectively identify and accurately read sEV-miRNAs in complex mixed samples. By integrating a material with low-background-signal and high-performance material with the original branch-strand migration logic gate system, we achieved an intelligent sensing platform that integrates capture, recognition, detection, and interpretation of sEV-miRNAs. We successfully demonstrated the response of the logic gate module of sEV-miRNAs in clinical samples, with detection results maintaining high consistency with conventional qRT-PCR methods. Our DNA logic gate platform boasts a simple design, adaptability, expandable complexity, and compatibility with various systems, enabling diverse possibilities for the nucleic acid-based development of intricate transduction networks in biomedical fields. This platform paves the way for more effective diagnosis, treatment, and monitoring of diseases, marking a significant advancement in nucleic acid signal amplification technologies and their utilization in biological computing.
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