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基于适体互补链杂交体和核酸外切酶 I 的拱形结构的新型电化学生物适体传感器用于灵敏检测链霉素。

A novel electrochemical aptasensor based on arch-shape structure of aptamer-complimentary strand conjugate and exonuclease I for sensitive detection of streptomycin.

机构信息

Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Research Institute of Sciences and New Technology, Mashhad, Iran.

Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.

出版信息

Biosens Bioelectron. 2016 Jan 15;75:123-8. doi: 10.1016/j.bios.2015.08.017. Epub 2015 Aug 11.

DOI:10.1016/j.bios.2015.08.017
PMID:26302000
Abstract

Detection and quantitation of antibiotic residues in blood serum and animal foodstuffs are of great significance. In this study, an electrochemical aptasensor was developed for sensitive and selective detection of streptomycin, based on exonuclease I (Exo I), complimentary strand of aptamer (CS), Arch-shape structure of aptamer (Apt)-CS conjugate and gold electrode. The designed aptasensor inherits characteristics of gold including large surface area and high electrochemical conductivity, as well as high sensitivity and selectivity of aptamer toward its target, property of Arch-shape structure of Apt-CS conjugate to act as a gate and barrier for the access of redox probe to the surface of electrode and the function of Exo I as an enzyme which selectively digests the 3'-end of single stranded DNA (ssDNA). In the absence of streptomycin the gate remains closed. Thus, the electrochemical signal is weak. Upon addition of streptomycin, the Apt leaves the CS and binds to streptomycin and the Arch-shape structure is disassembled. Then, Exo I addition leads to a strong electrochemical signal. The designed electrochemical aptasensor exhibited high selectivity toward streptomycin with a limit of detection (LOD) as low as 11.4nM. Moreover, the developed electrochemical aptasensor was successfully used to detect streptomycin in milk and serum with LODs of 14.1 and 15.3nM, respectively.

摘要

检测和定量血清和动物食品中的抗生素残留具有重要意义。本研究基于外切酶 I(Exo I)、适体互补链(CS)、适体 CS 缀合物的拱型结构和金电极,开发了一种用于灵敏和选择性检测链霉素的电化学适体传感器。设计的适体传感器继承了金的特性,包括大的表面积和高的电化学导电性,以及适体对其靶标具有的高灵敏度和选择性,适体 CS 缀合物的拱型结构作为门和障碍的特性,可防止氧化还原探针进入电极表面,以及 Exo I 作为酶的功能,可选择性地消化单链 DNA(ssDNA)的 3'-端。在没有链霉素的情况下,门保持关闭。因此,电化学信号很弱。加入链霉素后,适体离开 CS 并与链霉素结合,拱型结构被拆开。然后,加入 Exo I 会导致强烈的电化学信号。设计的电化学适体传感器对链霉素具有高选择性,检测限(LOD)低至 11.4nM。此外,该电化学适体传感器成功用于检测牛奶和血清中的链霉素,检测限分别为 14.1 和 15.3nM。

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