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基于金@银核壳纳米粒子的表面增强拉曼光谱杂交链式反应扩增法测定 17β-雌二醇

Determination of 17β-estradiol by surface-enhanced Raman spectroscopy merged with hybridization chain reaction amplification on Au@Ag core-shell nanoparticles.

机构信息

School of Food Science and Biological Engineering, Engineering Research Center of Bio-process, MOE, Hefei University of Technology, Hefei, 230009, China.

Hubei Key Laboratory of Edible Wild Plants Conservation and Utilization, Hubei Normal University, Huangshi, 435002, China.

出版信息

Mikrochim Acta. 2019 Jan 7;186(2):52. doi: 10.1007/s00604-018-3114-x.

Abstract

The authors describe an aptamer-based assay for 17β-estradiol. It relies on the combined use of surface enhanced Raman scattering (SERS) and hybridization chain reaction (HCR). The aptamer against 17β-estradiol is applied as the recognition probes, and this results in excellent specificity. Specific recognition of target 17β-estradiol induce the freedom of DNA 2, which will open the stem-loop structure of probe 1 on the Au@Ag and form the partial dsDNA structure. With the nicking enzyme, the partial dsDNA will be hydrolyzed and the reside ssDNA on Au@Ag will form a small stem-loop structure. With the help of the other probe 2 modified Au@Ag and pre-immobilized probe 3 on the well of the microplate, an enzyme-free HCR can occur and tremendous Au@Ag can be assembled along the formed dsDNA in HCR, which can act as the excellent substrate for Raman measurement and greatly amplify the Raman signal of R6G on the Au@Ag. Afterwards, the key factor, ratio between probe 2-Au@Ag (P2) and probe1-Au@Ag (P1), affects the detection sensitivity is systematically optimized for the best sensing performance. The SERS signal of R6G, best measured at 1651 cm, increases linearly in the wide range from 1 pM to 10 nM. The detection limit can be as low as 0.1 pM. Graphical abstract Schematic presentation of an aptamer-based surface enhanced Raman scattering method for accurate detection of 17β-estradiol, which is integrated with hybridization chain reaction for signal amplification and sensitivity improvement.

摘要

作者描述了一种基于适体的 17β-雌二醇分析方法。该方法依赖于表面增强拉曼散射(SERS)和杂交链式反应(HCR)的联合使用。针对 17β-雌二醇的适体被用作识别探针,从而实现了出色的特异性。目标 17β-雌二醇的特异性识别诱导 DNA 2 的自由,这将打开 Au@Ag 上探针 1 的茎环结构并形成部分双链 DNA 结构。带有切口酶,部分双链 DNA 将被水解,并且在 Au@Ag 上的残留 ssDNA 将形成小茎环结构。在另一个修饰有 Au@Ag 的探针 2 和预先固定在微孔板孔中的探针 3 的帮助下,可以发生无酶 HCR,并且可以沿着 HCR 中形成的 dsDNA 组装大量的 Au@Ag,这可以作为拉曼测量的极好底物并大大放大 Au@Ag 上 R6G 的拉曼信号。之后,系统优化了关键因素,即探针 2-Au@Ag(P2)和探针 1-Au@Ag(P1)之间的比例,以获得最佳的传感性能。R6G 的 SERS 信号在 1651 cm 处最佳测量,在 1 pM 至 10 nM 的宽范围内呈线性增加。检测限可以低至 0.1 pM。摘要 基于适体的表面增强拉曼散射方法示意图,用于准确检测 17β-雌二醇,该方法与杂交链式反应集成在一起,用于信号放大和灵敏度提高。

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