在这项研究中，展示了基于双感知/作用途径的新型感知策略，能够触发基于DNA的AND门，用于针对乙型肝炎病毒DNA（HBV-DNA）的敏感、选择性检测。这种方法涵盖了一个逻辑操作的酶机制，其操作基于生理相关生物标志物的变化，用于检测肝功能损伤。丙氨酸转移酶（ALT）和乳酸脱氢酶（LDH）被用作AND门的输入，生成一个输出信号——乳酸。特别地，乳酸在阳极电极处被与介导电子转移的费洛甲烷基团相连的乳酸氧化酶氧化回到丙酮酸（创造了一个放大的循环机制）。阳极电极进一步连接着MvBOx基于Myrothecium verrucaria的胆红素氧化酶（MvBOx）基生物阴极，该阴极经SiO2纳米颗粒（SiO2NPs）功能化的苯硼酸和三甲基咖啡碱改性，触发了电极表面局部pH变化时的猝灭DNA（qDNA）释放（值得注意的是，改性的SiONPs在局部pH梯度下得到负电荷释放负电荷的DNA)。接下来，标记着BHQ2和检测DNA（标记着FAM的dDNA）的释放的qDNA检测到了HBV-DNA。所提出的生物传感器能够区分缓冲溶液和人血清中HBV-DNA存在和不存在的阈值，分别为0.05 fM和1 fM。这种酶/DNA逻辑网络对未来的生物医学应用（例如早期检测肝癌等疾病）非常有价值。在未来的技术发展中，这种技术可以很容易地与智能手机相机集成，实现更为用户友好的应用。版权所有©2023 Elsevier B.V.。保留所有权利。

In this study, a novel sensing strategy based on double sensing/actuating pathway is demonstrated, being capable to trigger the DNA-based AND gate for the sensitive and selective detection of hepatitis B virus DNA (HBV-DNA). Such an approach encompasses an enzymatic machinery logically operated using the variation of physiologically relevant biomarkers for liver dysfunctions. Alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) are used as inputs of an AND gate generating an output signal, namely lactate. In particular, lactate is oxidized back to pyruvate at the anodic electrode by lactate oxidase connected in mediated electron transfer through ferrocene moieties (creating an amplifying recycling mechanism). The anodic electrode is further connected with a Myrothecium verrucaria bilirubin oxidase (MvBOx) based biocathode modified with SiO2 nanoparticles (SiO2NPs) functionalized with phenyl boronic acid and trigonelline, triggering the release of quenching DNA (qDNA) upon local pH change at the electrode surface (notably, modified SiONPs gets negatively recharged upon local pH gradient releasing negatively charged DNA). Next, the released qDNA labeled with BHQ2 and detecting DNA (dDNA, labeled with FAM) are detecting HBV-DNA. The proposed biosensor can discriminate between the absence and presence of HBV-DNA setting the threshold at 0.05 fM in model buffer solutions and 1 fM in human serum. This enzymatic/DNA logic network can be of particular interest for future biomedical applications (e.g., early detection of liver cancer disease etc.). In the future development this technology could be easily integrated with a smartphone camera, allowing more user-friendly applications.Copyright © 2023 Elsevier B.V. All rights reserved.