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基于 T-Hg-T 触发杂交链式反应扩增策略的金纳米粒子修饰生物传感器用于汞检测的蒲公英状氧化铜微球的设计

Dandelion-like CuO microspheres decorated with Au nanoparticle modified biosensor for Hg detection using a T-Hg-T triggered hybridization chain reaction amplification strategy.

机构信息

School of Public Health and Management, Chongqing Medical University, Chongqing, China; Joint International Research Laboratory of Reproduction & Development, Chongqing Medical University, Chongqing, China.

College of Pharmacy, Chongqing Medical University, Chongqing, China.

出版信息

Biosens Bioelectron. 2019 Apr 15;131:207-213. doi: 10.1016/j.bios.2019.01.063. Epub 2019 Feb 19.

Abstract

We fabricate a novel electrochemical biosensor based on the specific thymine-Hg-thymine (T-Hg-T) base pair for the highly sensitive detection of mercury ions (Hg) and utilize toluidine blue (TB) as a redox indicator that is combined with a hybridization chain reaction (HCR) for signal amplification. The dandelion-like CuO (D-CuO) microspheres that were assembled using Au nanoparticles were first introduced as support materials, which produced more active sites for the thiolated probe (P1) combination. Then, the presence of Hg induced P1 to hybridize with the other oligonucleotide (P2) through Hg-mediated T-Hg-T complexes. In addition, the partial sequence of P2 acted as an initiator sequence, which led the two hairpin DNA (H1 and H2) strands to collectively form the extended double-strand DNA through the HCR process on the electrode surface. TB was employed to interact with the double strands and produce an efficient electrochemical signal. The proposed strategy combined the amplification of the HCR and the inherent redox activity of TB and utilized D-CuO/Au composites, which exhibited high sensitivity for Hg determination. Under the optimum conditions, the proposed biosensor showed a prominent response for Hg, including a linear range from 1 pM to 100 nM and a detection limit of 0.2 pM (S/N = 3). Moreover, the new biosensor proved its potential application for trace Hg determination in environmental water samples.

摘要

我们基于胸腺嘧啶 - Hg - 胸腺嘧啶(T-Hg-T)特异碱基对,制造了一种新型电化学生物传感器,用于高灵敏度检测汞离子(Hg),并利用甲苯胺蓝(TB)作为与杂交链式反应(HCR)结合的氧化还原指示剂,用于信号放大。首先,将组装有金纳米粒子的蒲公英状氧化铜(D-CuO)微球用作支撑材料,这为巯基探针(P1)的组合产生了更多的活性位点。然后,Hg 的存在通过 Hg 介导的 T-Hg-T 复合物诱导 P1 与另一个寡核苷酸(P2)杂交。此外,P2 的部分序列充当起始序列,使得两条发夹 DNA(H1 和 H2)链通过在电极表面上的 HCR 过程共同形成扩展的双链 DNA。TB 用于与双链相互作用并产生有效的电化学信号。所提出的策略结合了 HCR 的放大和 TB 的固有氧化还原活性,并利用 D-CuO/Au 复合材料,对 Hg 测定表现出高灵敏度。在最佳条件下,所提出的生物传感器对 Hg 表现出显著的响应,包括 1 pM 至 100 nM 的线性范围和 0.2 pM(S/N = 3)的检测限。此外,新的生物传感器证明了其在环境水样中痕量 Hg 测定中的潜在应用。

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