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

Journal 2023 · IF 6.2 · 发表于 2023 年 3 月 8 日
作者: Xu Wen,Xueling Yan,Xiaoling Zheng,Qiming Kou,Lulan Yang,Jiaming Tang,Xingyue Chen,Yong Xie,Tao Le
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摘要

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 d  = 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.

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