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Visual detection of lead(II) using a label-free DNA-based sensor and its immobilization within a monolithic hydrogel

机译:使用无标记的基于DNA的传感器对铅(II)进行视觉检测并将其固定在整体水凝胶中

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摘要

Lead is highly toxic and its detection has attracted a lot of research interests. In recent years, DNA has been used for Pb2+ recognition and many fluorescent sensors with low to sub-nM detection limits have been reported. These figures of merit were typically measured using a spectrophotometer that can detect nM DNA with a high signal-to-noise ratio. For visual detection, however, μM DNA or dye was required, making it difficult to detect low nM targets. We recently achieved a visual sensitivity of 10 nM Hg2+ by immobilizing a DNA probe in a hydrogel. This was made possible because the gel was able to actively adsorb Hg2+. In this work, we aim to test whether this method can be extended to the detection of Pb2+. First, a new Pb2+ sensor was designed based on a guanine-rich DNA and DNA binding dyes such as thiazole orange and SYBR Green I. The free DNA showed a detection limit of 8 nM Pb2+ using 40 nM DNA. For visual detection in solution with 1 μM of the DNA probe, however, ∼300 nM Pb2+ was required. After immobilization in a monolithic polyacrylamide hydrogel, even 20 nM Pb2+ could be visually detected with a sample volume of 50 mL. Therefore, sensitive detection without signal amplification was achieved. Finally, we demonstrated simultaneous detection of both Hg2+ and Pb2+ in the same water sample with shape encoded hydrogel sensors.
机译:铅具有剧毒,其检测吸引了许多研究兴趣。近年来,DNA已被用于Pb2 +的识别,并且已经报道了许多低至亚nM检出限的荧光传感器。这些品质因数通常是使用能检测高信噪比的nM DNA的分光光度计测量的。但是,为了进行视觉检测,需要使用μMDNA或染料,这使得难以检测低nM靶标。我们最近通过将DNA探针固定在水凝胶中实现了10 nM Hg2 +的视觉灵敏度。由于凝胶能够主动吸附Hg2 +,因此使之成为可能。在这项工作中,我们旨在测试该方法是否可以扩展到检测Pb2 +。首先,基于富含鸟嘌呤的DNA和DNA结合染料(如噻唑橙和SYBR Green I)设计了一种新的Pb2 +传感器。使用40 nM DNA时,游离DNA的检测限为8 nM Pb2 +。但是,要在含有1μMDNA探针的溶液中进行视觉检测,需要约300 nM Pb2 +。固定在整体聚丙烯酰胺水凝胶中后,即使样品体积为50 mL,也可以目视检测到20 nM Pb2 +。因此,实现了无需信号放大的灵敏检测。最后,我们演示了使用形状编码水凝胶传感器同时检测同一水样中的Hg2 +和Pb2 +。

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