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基于包含较少涉及适配体碱基对的稳定 DNA 双链探针的快速简单的比率荧光传感器用于棒曲霉素检测。

A rapid and simple ratiometric fluorescent sensor for patulin detection based on a stabilized DNA duplex probe containing less amount of aptamer-involved base pairs.

机构信息

Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

Research Institute of Sciences and New Technology, Mashhad, Iran.

出版信息

Talanta. 2019 Nov 1;204:641-646. doi: 10.1016/j.talanta.2019.06.057. Epub 2019 Jun 15.

Abstract

In this study, a sensor is described for determination of patulin by using ratiometric fluorescence measurement and strand displacement strategy. In the presence of patulin, the ratiometric fluorescence response decreases, owing to disassembly of DNA duplex structure and target-mediated release of TAMRA-labeled complementary DNA sequence2 (cDNA2). While, in the absence of target, the fluorescence resonance energy transfer (FRET) phenomenon happens between FAM and TAMRA under excitation at 490 nm, resulting in the enhancement of ratiometric signal. The use of ratiometric fluorescence signal with different signal indicators avoids the problem of environmental interference and improves the sensitivity of the aptasensor. Also, the DNA duplex structure contains minimum aptamer-involved base pair sequence, resulting in further improvement of the aptasensor sensitivity. This sensing platform provided a wide linear range from 15 ng/L to 35 μg/L and a detection limit of 6 ng/L for patulin. The aptasensor was used to determine patulin in spiked apple juice samples and showed satisfactory results.

摘要

本研究描述了一种基于比率荧光测量和链置换策略的传感器,用于测定棒曲霉素。在棒曲霉素存在的情况下,由于 DNA 双链结构的解体和靶介导的 TAMRA 标记互补 DNA 序列 2(cDNA2)的释放,比率荧光响应降低。而在没有靶标时,在 490nm 激发下,FAM 和 TAMRA 之间发生荧光共振能量转移(FRET)现象,导致比率信号增强。使用具有不同信号指示剂的比率荧光信号可避免环境干扰问题,并提高适体传感器的灵敏度。此外,DNA 双链结构包含最小的适体涉及碱基对序列,从而进一步提高了适体传感器的灵敏度。该传感平台为棒曲霉素提供了从 15ng/L 到 35μg/L 的宽线性范围和 6ng/L 的检测限。该适体传感器用于测定加标苹果汁样品中的棒曲霉素,结果令人满意。

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