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<p>Aptamer-based detection methods have emerged as a promising research direction for detecting gentamicin (GEN). Herein, the aptamer for GEN with <i>K</i><sub>d</sub> of 4.8 µM was firstly selected by non-immobilized graphene oxide (GO)-SELEX. With the molecular docking simulations and truncation strategies, the binding affinity of the truncated aptamer GEN-T-1 (53 nt) with <i>K</i><sub>d</sub> of 0.4 µM was further improved. Based on this, a colorimetric aptasensor for GEN detection was developed with GEN-T-1 as the recognition element and AuNPs@CeO<sub>2</sub> as the nanozyme which could catalyze oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB). In presence of GEN, the aptamer selectively bound to GEN, dissociating from nanozyme surface and generating a lighter color. The aptasensor demonstrated a low limit of detection 3.56 ng/mL and a wide linear detection range of 10–10,000 ng/mL, with excellent specificity, stability, repeatability, and reproducibility. The method was successfully applied to detect GEN in milk with recoveries of 97.97%-100.49%. The research results provide a theoretical foundation for the rapid and accurate detection of GEN.</p> Graphical Abstract <p></p>

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GO-SELEX of aptamer for colorimetric aptasensor based on AuNPs@CeO2 nanozyme in detection of gentamicin residues in milk

  • 期刊论文

  • 发表日期：

    2026年5月9日

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  • Fengling Yue,
  • Miaomiao Sui,
  • Jian Liu,
  • Bo Wang,
  • Lijun Jiang,
  • Yifan Du,
  • Jiayi Ye,
  • Yingying Nie,
  • Guihong Zhao

摘要

Aptamer-based detection methods have emerged as a promising research direction for detecting gentamicin (GEN). Herein, the aptamer for GEN with Kd of 4.8 µM was firstly selected by non-immobilized graphene oxide (GO)-SELEX. With the molecular docking simulations and truncation strategies, the binding affinity of the truncated aptamer GEN-T-1 (53 nt) with Kd of 0.4 µM was further improved. Based on this, a colorimetric aptasensor for GEN detection was developed with GEN-T-1 as the recognition element and AuNPs@CeO2 as the nanozyme which could catalyze oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB). In presence of GEN, the aptamer selectively bound to GEN, dissociating from nanozyme surface and generating a lighter color. The aptasensor demonstrated a low limit of detection 3.56 ng/mL and a wide linear detection range of 10–10,000 ng/mL, with excellent specificity, stability, repeatability, and reproducibility. The method was successfully applied to detect GEN in milk with recoveries of 97.97%-100.49%. The research results provide a theoretical foundation for the rapid and accurate detection of GEN.

Graphical Abstract