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Highly Selective DNA-Based Sensor for Lead(II) and Mercury(II) Ions

机译:基于高选择性DNA的铅(II)和汞(II)离子传感器

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We have developed a technique for the highly selective and sensitive detection of Pb~(2+) and Hg~(2+) using a thrombin-binding aptamer (TBA) probe labeled with the donor carboxyfluorescein (FAM) and the quencher 4-([4-(dimethylamino)phenyl]azo)benzoic acid (DAB-CYL) at its 5' and 3' termini, respectively. The TBA has a random coil structure that changes into a G-quartet structure and a hairpin-like structure upon binding Pb~(2+) and Hg~(2+) ions, respectively. As a result, the fluorescence decreases through fluorescence resonance energy transfer (FRET) between the fluorophore and quencher. These changes in fluorescence intensity allow the selective detection of Pb~(2+) and Hg~(2+) ions at concentrations as low as 300 pM and 5.0 nM using this TBA probe in the presence of phytic acid and a random DNA/NaCN mixture, respectively. The linear correlation existed between the fluorescence intensity and the concentration of Pb~(2+) and Hg~(2+) over the range of 0.5-30 nM (R~(2) velence 0.98) and 10-200 nM (R~(2) velence 0.98), respectively. To the best of our knowledge, this is the first example of a single DNA-based sensor that allows the detection of both Hg~(2+) and Pb~(2+) ions. This simple and cost-effective probe was also applied to separately determine Pb~(2+) in soil samples and spiked Hg~(2+) in pond samples.
机译:我们已经开发了一种技术,该技术使用标记有供体羧基荧光素(FAM)和淬灭剂4-(-)的凝血酶结合适体(TBA)探针对Pb〜(2+)和Hg〜(2+)进行高选择性和灵敏的检测。 [4-(二甲基氨基)苯基]偶氮)苯甲酸(DAB-CYL)分别位于其5'和3'末端。该TBA具有无规卷曲结构,当结合Pb〜(2+)和Hg〜(2+)离子时,其变为G四重结构和发夹状结构。结果,荧光通过荧光团和猝灭剂之间的荧光共振能量转移(FRET)而减少。这些荧光强度的变化允许在存在植酸和随机DNA / NaCN的情况下使用该TBA探针选择性检测浓度低至300 pM和5.0 nM的Pb〜(2+)和Hg〜(2+)离子分别。荧光强度与Pb〜(2+)和Hg〜(2+)的浓度在0.5-30 nM(R〜(2)速度0.98)和10-200 nM(R〜 (2)velence 0.98)。据我们所知,这是单个基于DNA的传感器的第一个示例,该传感器可以检测Hg〜(2+)和Pb〜(2+)离子。该简单且具有成本效益的探针还用于分别测定土壤样品中的Pb〜(2+)和池塘样品中的加标Hg〜(2+)。

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