Aptamer-based fluorometric determination of ATP by using target-cycling strand displacement amplification and copper nanoclusters
Aptamer-based fluorometric determination of ATP by using target-cycling strand displacement amplification and copper nanoclusters
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使用靶循环链置换扩增和铜纳米簇进行基于适体的 ATP 荧光测定
DOI：
10.1007/s00604-017-2337-6
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发表时间：
2017-06
影响因子：
5.7
通讯作者：
Jiang Jian-Hui
中科院分区：
文献类型：
--
作者：
Wang Yu-Min;Liu Jin-Wen;Duan Lu-Ying;Liu Si-Jia;Jiang Jian-Hui
描述了一种荧光适体传感器，它结合了靶标循环链置换扩增 (TCSDA) 和以双链 DNA (dsDNA) 为模板的铜纳米簇 (CuNC) 的合成。具体来说，该检测方案通过靶标诱导的结构转换设计应用于 5'-三磷酸腺苷 (ATP) 的测定。适体发夹探针 (AP) 与 ATP 的结合诱导从发夹形状到开放构象的结构转变。这有利于与引物的杂交并触发 TCSDA 反应。该扩增步骤产生大量的 dsDNA，可以直接作为合成荧光 CuNC 的模板，从而产生强烈的红色荧光（激发/发射最大值为 340/598 nm），可用于定量 ATP。该检测的动态范围可延伸超过 4 个十年（从 0.01 nM 到 100 nM），检测限为 5 pM。可以想象，这种检测方案适用于许多其他有合适适配体的分析物。图形摘要 ATP 测定原理图（检测限为 5 pM），该测定利用了 (a) 靶标循环链置换扩增和 (b) 红色荧光铜纳米簇。 KF聚合酶：聚合酶的克列诺片段
A fluorescence aptasensor is described that combines target-cycling strand displacement amplification (TCSDA) and synthesis of copper nanoclusters (CuNCs) templated with double-stranded DNA (dsDNA). Specifically, the detection scheme was applied to the determination of adenosine-5′-triphosphate (ATP) via target-induced structure switching design. The binding of an aptamer hairpin probe (AP) to ATP induces a structural switch from a hairpin shape to an open conformation. This facilitates hybridization with a primer and triggers a TCSDA reaction. This amplification step produces a large quantity of dsDNA that can directly act as a template for the synthesis of fluorescent CuNCs, thereby producing a strong red fluorescence (with excitation/emission maxima at 340/598 nm) that can be used to quantify ATP. The assay has a dynamic range that extends over 4 decades (from 0.01 nM to 100 nM) and a 5 pM detection limit. Conceivably, this detection scheme is applicable to numerous other analytes for which suitable aptamers are available.Graphical abstractSchematic of an ATP assay (with a 5 pM detection limit) that is taking advantages of (a) target-cycling strand displacement amplification, and (b) of red fluorescent copper nanoclusters. KF polymerase: Klenow fragment of polymerase
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