We developed a simple and selective fluorescent aptasensor for the detection of oxytetracycline (OTC) using aptamer-conjugated gold nanoparticles (AuNPs) and a complex formed between oxytetracycline and a europium cation. In this study, AuNPs were modified with an OTC aptamer using two methods (Tween and NaCl methods). In the absence of OTC, an OTC-Eu3+ complex binds to the aptamer in the aptamer-conjugated AuNPs to give weak fluorescence emission. However, in the presence of OTC, the aptamer interacts with its target, causing a strong fluorescence emission. Under optimum conditions, the designed method showed high selectivity for OTC and a good linear range to OTC concentration from 15 to 500 nM with a limit of detection (LOD) of 10.6 nM for the NaCl method and linear range over 15–500 nM with an LOD of 8.8 nM for the Tween method. This biosensor was successfully employed to quantify OTC in milk and tablet samples.

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