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适体的筛选和截断作为荧光传感平台,用于选择性和灵敏检测呋喃西林。

Selection and truncation of aptamers as fluorescence sensing platforms for selective and sensitive detection of nitrofurazone.

机构信息

College of Life Science, Chongqing Normal University, Chongqing, 401331, PR China.

Bioassay 3D Reconstruction Laboratory, Chongqing College of Electronic Engineering, Chongqing, 401331, PR China.

出版信息

Anal Chim Acta. 2023 Apr 29;1252:341044. doi: 10.1016/j.aca.2023.341044. Epub 2023 Mar 8.

Abstract

Nitrofurazone (NFZ) is an antibiotic banned in many countries, as its residue seriously harms the human body. Herein, anti-NFZ aptamers were selected and identified based on the magnetic bead SELEX technique using a ssDNA library with a full length of 90 nucleotides (nt). Five full sequence candidate aptamers (NFZ8, NFZ24, NFZ28, NFZ34, and NFZ70) were obtained by secondary structure analysis. We optimized the entire sequence to obtain a truncated aptamer, a 16 nt sequence (NFZ8-1:5'-GTTCTATTGAAAAAAC-3') that showed the highest affinity for NFZ (K = 76.11 nM). The binding site of NFZ and aptamer NFZ8-1 was found to be "GAA" by molecular docking. In addition, utilizing the most special truncated aptamer NFZ8-1 as the identification probe, a graphene oxide fluorescent aptasensor is an innovative for the detection of NFZ residue that showed a wide linear reach from 1.25 to 160 ng/mL and a low limit of detection of 1.13 ng/mL. In the actual water environment sample detection, the recovery rate ranged from 95.21 to 113.66%, and the coefficient of variation ranged from 3.53 to 11.24%. These results demonstrate that the NFZ-truncated aptamer applied to the aptasensor provides a novel methodology for recognizing NFZ residues.

摘要

硝呋太尔(NFZ)是一种在许多国家被禁用的抗生素,因为其残留会严重危害人体健康。本研究采用磁珠 SELEX 技术,以全长为 90 个核苷酸(nt)的 ssDNA 文库为基础,筛选并鉴定了抗 NFZ 的适体。通过二级结构分析,获得了 5 条全长候选适体(NFZ8、NFZ24、NFZ28、NFZ34 和 NFZ70)。我们对整个序列进行了优化,得到了一个截断的适体,一个 16 个核苷酸序列(NFZ8-1:5'-GTTCTATTGAAAAAAC-3'),对 NFZ 表现出最高的亲和力(K=76.11 nM)。通过分子对接发现,NFZ 与适体 NFZ8-1 的结合位点为“GAA”。此外,利用最特殊的截断适体 NFZ8-1 作为识别探针,构建了一种基于石墨烯氧化物荧光适体传感器的新型 NFZ 残留检测方法,其线性范围从 1.25 到 160 ng/mL,检测限低至 1.13 ng/mL。在实际水环境样品检测中,回收率范围为 95.21%至 113.66%,变异系数范围为 3.53%至 11.24%。这些结果表明,应用于适体传感器的 NFZ 截断适体为识别 NFZ 残留提供了一种新方法。

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