A label-free Exonuclease I-assisted fluorescence aptasensor for highly selective and sensitive detection of silver...

Based on the specific interaction of Ag⁺ and cytosine-cytosine (C-C) base mismatch and using berberine (Ber) as the fluorescent probe and Exonuclease I (Exo I) as the background fluorescence reducing tool, a label-free Exo I-assisted fluorescence aptamer sensing platform was established for the detection of silver ions with high sensitivity and selectivity. Exo I reduced the fluorescence background of the Ber/Ag⁺-aptamer complex to a level similar to that of Ber itself in the absence of Ag⁺. After introducing Ag⁺ into the sensing system, it induces the aptamer rich in base C to form C-Ag⁺-C i-motif structure which are resistant to degradation mediated by Exo I. The concentration of Ber, Ag⁺-aptamer, Exo I and the temperature and reaction time for Exo I were all optimized. Under the optimal experimental conditions, the detection limit of Ag⁺ was 4.4 nM and the linear range was from 0.0059 μM to 235.48 μM with a coefficient of determination (R²) > 0.99. Moreover, the proposed strategy had been successfully applied to the detection of Ag⁺ in tap water and human serum with a good recovery ranging from 88.4% to 106.9%.