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基于特异性结构胸腺嘧啶-汞离子-胸腺嘧啶的汞离子无标记共振光散射检测

[Label-Free Resonance Light Scattering Detection of Hg²⁺ Based on Specific Structure Thymine-Hg²⁺-Thymine].

作者信息

Yang Sheng-yuan, Xu Xiao-na, Yu Jun-hui, Yang Hui-xian, Hu Cheng-lan

出版信息

Guang Pu Xue Yu Guang Pu Fen Xi. 2015 Dec;35(12):3471-4.

Abstract

A new label-free resonance light scattering method for the highly selective and sensitive detection of mercury ion was designed. This strategy makes use of the target-induced DNA conformational change to enhance the resonance light scattering intensity leading to an amplified optical signal. The Hg²⁺ ion, which possesses a unique property to bind specifically to two DNA thymine (T) bases, in the presence of Hg²⁺, the specific oligonucleotide probes form a conformational reorganization of the oligonucleotide probes from single-chain structure to duplex-like complexes, which can greatly enhance the resonance light scattering intensity. Under the optimum experimental conditions, the enhanced resonance light scattering intensity at 566 nm was in proportion of mercury ion concentration in the range 7.2 x 10⁻⁹ x 10⁻⁸ mol · L⁻¹ with the linear regression equation was ΔI = 5.12c+3.55 (r = 0.999 5). This method was successfully applied to detection of Hg²⁺ in enviro nmental water samples, the RSD were less than 1.9% and recoveries were 99.4%-104.3%. This label-free strategy uses the mercury specific oligonucleotide probes as recognition elements and control the strength of resonance light scattering by changing the concentration of Hg²⁺. It translating the small molecule detection into the DNA hybridization behavior leading to an amplified resonance light scattering signal can well enhance the sensitive detection of Hg²⁺. With amplification by DNA hybridization behavior, the sensitivity for the detection of Hg²⁺ can achieve 2.16 x 10⁻⁹ mol · L(⁻¹). In this study, the stacked T-Hg²⁺-Tfunctioned not only as amplification property but also as an selective recognition. The highly specific detection of Hg²⁺ is attributed to the formation of a stable T-Hg²⁺-T complex.

摘要

设计了一种用于高选择性和灵敏检测汞离子的新型无标记共振光散射方法。该策略利用目标诱导的DNA构象变化来增强共振光散射强度,从而产生放大的光信号。Hg²⁺离子具有与两个DNA胸腺嘧啶(T)碱基特异性结合的独特性质,在Hg²⁺存在下,特异性寡核苷酸探针形成从单链结构到双链样复合物的寡核苷酸探针构象重组,这可以大大增强共振光散射强度。在最佳实验条件下,566 nm处增强的共振光散射强度与汞离子浓度在7.2×10⁻⁹~10⁻⁸mol·L⁻¹范围内成正比,线性回归方程为ΔI = 5.12c + 3.55(r = 0.999 5)。该方法成功应用于环境水样中Hg²⁺的检测,相对标准偏差小于1.9%,回收率为99.4%~104.3%。这种无标记策略使用汞特异性寡核苷酸探针作为识别元件,并通过改变Hg²⁺的浓度来控制共振光散射的强度。它将小分子检测转化为DNA杂交行为,导致放大的共振光散射信号,能够很好地增强Hg²⁺的灵敏检测。通过DNA杂交行为进行放大,Hg²⁺检测的灵敏度可达到2.16×10⁻⁹mol·L⁻¹。在本研究中,堆积的T-Hg²⁺-T不仅起到放大作用,还起到选择性识别作用。Hg²⁺的高特异性检测归因于稳定的T-Hg²⁺-T复合物的形成。

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