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新发现的硫黄素T与DNA G-四链体的竞争性结合以及一种用于尿液样本中钾离子测定的具有超低检测限的无标记荧光生物传感器的开发。

Newly found K-Thioflavin T competitive binding to DNA G-quadruplexes and the development of a label-free fluorescent biosensor with extra low detection limit for K determination in urine samples.

作者信息

Chitbankluai Khwanrudee, Thavarungkul Panote, Kanatharana Proespichaya, Kaewpet Morakot, Buranachai Chittanon

机构信息

Division of Physical Science, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90110, Thailand; Center of Excellence for Trace Analysis and Biosensor, Prince of Songkla University, Hat Yai, Songkhla, 90110, Thailand; Thailand Center of Excellence in Physics, Commission on Higher Education, 328 Si Ayutthaya Road, Bangkok, 10400, Thailand.

Division of Physical Science, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90110, Thailand; Center of Excellence for Trace Analysis and Biosensor, Prince of Songkla University, Hat Yai, Songkhla, 90110, Thailand; Thailand Center of Excellence in Physics, Commission on Higher Education, 328 Si Ayutthaya Road, Bangkok, 10400, Thailand; Center of Excellence for Innovation in Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90110, Thailand.

出版信息

Spectrochim Acta A Mol Biomol Spectrosc. 2022 Aug 5;276:121244. doi: 10.1016/j.saa.2022.121244. Epub 2022 Apr 8.

Abstract

The determination of potassium ion K in body fluids is important in health monitoring and diagnoses. One of the interesting and simple methods for K detection is the use of label-free biosensors based on DNA G-quadruplexes (GQs) coupled with a specific fluorescent probe, such as Thioflavin T (ThT), which lights up when bound with K-stabilized GQs. However, these biosensors are not generally sensitive. In this work, we found a solution: at a low concentration, K competes with ThT in binding to a bimolecular GQ or a tetramolecular GQ, resulting in a decrease in ThT fluorescence emission with increasing K. Therefore, we developed a label-free turn-off fluorescent K sensor. The sensor provides a very low detection limit of 21.87 ± 0.59 nM. Other possible interfering components in urine did not exert any effect even at quantities that were 10-fold greater than their upper limit of normal concentrations found in urine samples. With its only requirement of diluting samples, the developed low-cost label-free probe and simple sensor was successfully applied to the direct detection of K in normal urine samples with high accuracy (recoveries ranged from 90% to 100%).

摘要

测定体液中的钾离子K对健康监测和诊断很重要。一种有趣且简单的钾离子检测方法是使用基于DNA G-四链体(GQs)并结合特定荧光探针(如硫黄素T(ThT))的无标记生物传感器,当ThT与钾离子稳定的GQs结合时会发光。然而,这些生物传感器通常灵敏度不高。在这项工作中,我们找到了解决方案:在低浓度下,钾离子与硫黄素T竞争结合双分子GQ或四分子GQ,导致随着钾离子浓度增加硫黄素T荧光发射减弱。因此,我们开发了一种无标记的关闭型荧光钾离子传感器。该传感器的检测限低至21.87±0.59 nM。尿液中其他可能的干扰成分即使含量比尿液样本中正常浓度上限高10倍也不会产生任何影响。由于只需对样品进行稀释,所开发的低成本无标记探针和简单传感器成功应用于直接高精度检测正常尿液样本中的钾离子(回收率在90%至100%之间)。

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