AbstractA colorimetric assay for ATP is described that uses a strategy that combines the concept of split Mg(II)-dependent DNAzyme, split aptamer, and hybridization-induced aggregation of gold nanoparticles (AuNPs). Both ATP aptamer and Mg(II)-dependent DNAzyme are split into two fragments which are allocated to two well-designed DNA probes. The probes also possess mutually complementary stem sequences and spacer sequences. In the presence of ATP, the separated DNAzyme sequences in the two probes assemble via the synchronous recognition of ATP with two fragments of the aptamer. Then, the activated DNAzyme catalyzes multiple cycles of the cleavage of its substrate DNA sequence. The latter acts as a linker and induces the aggregation of two types of ssDNA-modified AuNP through the hybridization between the complementary sequences. Thus, the color of the AuNP solution remains red. However, in the absence of ATP, the detached aptamer cannot induce the assembly of DNAzyme to cleave the linker DNA. This results in the aggregation of AuNP and a concomitant color transition from red to purple. This ATP assay, performed at a wavelength of 530 nm, has a linear detection range that extends from 10 pM to 100 nM, with a detection limit of 5.3 pM. It was applied to the detection of ATP in human serum. Conceivably, the strategy has a wide scope in that it may be applied to the colorimetric detection of various other analytes through the split aptamer configuration. Graphical abstractSchematic presentation of colorimetric assay for adenosine triphosphate (ATP) based on the use of a split Mg(II)-dependent DNAzyme, a split aptamer, and by exploiting the hybridization-induced aggregation of gold nanoparticles that leads to a color change from red to purple

中文翻译：

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基于使用镁 (II) 依赖性 DNAzyme 的比色 ATP 测定

摘要描述了一种 ATP 比色测定，它使用一种策略，该策略结合了分裂 Mg(II) 依赖性 DNAzyme、分裂适体和杂交诱导的金纳米粒子 (AuNPs) 聚集的概念。ATP 适体和 Mg(II) 依赖性 DNAzyme 被分成两个片段，这些片段分配给两个精心设计的 DNA 探针。探针还具有相互互补的茎序列和间隔序列。在存在 ATP 的情况下，两个探针中分离的 DNAzyme 序列通过 ATP 与两个适体片段的同步识别组装。然后，活化的 DNAzyme 催化其底物 DNA 序列切割的多个循环。后者充当接头并通过互补序列之间的杂交诱导两种类型的 ssDNA 修饰的 AuNP 的聚集。因此，AuNP 溶液的颜色保持红色。然而，在没有 ATP 的情况下，分离的适体不能诱导 DNAzyme 的组装以切割接头 DNA。这导致 AuNP 的聚集和伴随的从红色到紫色的颜色转变。这种 ATP 检测在 530 nm 的波长下进行，具有从 10 pM 到 100 nM 的线性检测范围，检测限为 5.3 pM。应用于人血清中ATP的检测。可以想象，该策略具有广泛的范围，因为它可以通过拆分适体配置应用于各种其他分析物的比色检测。图形摘要基于使用分裂 Mg(II) 依赖性 DNAzyme、分裂适体、三磷酸腺苷 (ATP) 比色测定的示意图，