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A novel enzyme-free and label-free fluorescence aptasensor for amplified detection of adenosine

机译:一种新型的无酶,无标记的荧光适体传感器,用于腺苷的扩增检测

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摘要

A novel enzyme-free and label-free fluorescence aptasensor based on target-catalyzed hairpin self-assembly is developed for amplified detection of adenosine. This aptasensor contains four DNA strands termed as aptamer-catalysis strand, inhibit strand, hairpin structures H1 and H2 which are partially complementary. Meanwhile, a sequence that can form DNA G-quadruplex is partly hidden in the stem of H2. In the absence of adenosine, aptamer-catalysis strand is inhibited, and cannot trigger the self-assembly between H1 and H2. Upon the addition of adenosine, the binding event of aptamer and adenosine triggers the self-assembly between H1 and H2, resulting in the formation of G-quadruplex at the end of H1-H2 complex. The addition of N-methyl mesoporphyrin IX, which has a pronounced structural selectivity for G-quadruplex, generates label-free fluorescence signal. In the optimum conditions, we could detect adenosine as low as 6 μM by monitoring the change in fluorescence intensity. Furthermore, this amplified aptasensor shows high selectivity toward adenosine against its analogs due to the specific recognition ability of the aptamer for the target. Thus, the proposed aptasensor could be used as a simple and selective platform for target detection.
机译:新型的基于目标催化的发夹自组装的无酶,无标记的荧光适体传感器被开发用于腺苷的扩增检测。该适体传感器包含四个DNA链,称为适体催化链,抑制部分互补的发夹结构H1和H2。同时,可以形成DNA G-四链体的序列部分隐藏在H2的茎中。在没有腺苷的情况下,适体催化链被抑制,并且不能触发H1和H2之间的自组装。加入腺苷后,适体和腺苷的结合事件触发了H1和H2之间的自组装,导致在H1-H2复合物末端形成G-四链体。 N-甲基中卟啉IX的添加对G-四链体具有明显的结构选择性,可生成无标记的荧光信号。在最佳条件下,我们可以通过监测荧光强度的变化来检测低至6μM的腺苷。此外,由于适体对靶标的特异性识别能力,该扩增的适体传感器对腺苷对其类似物显示出高选择性。因此,提出的适体传感器可以用作目标检测的简单和选择性平台。

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