A simple and label-free fluorescent DNA sensor for visual detection of aptamer-based berberine

Abstract

The intrinsic fluorescence of berberine is very weak, which can be enhanced by its interaction with specific aptamers. A simple and sensitive DNA sensor for visual detection of berberine has here been established. When using this sensor, there was a good linear relationship between the change in fluorescence intensity of berberine and the concentration of berberine in the range of 16–2000 nM, with a detection limit of 5.1 nM. The change in fluorescence intensity was caused by the addition of aptamers. A detection limit of 170.1 nM was acquired by reading the RGB values of fluorescent images with a smartphone for the quantification of berberine. Common antibiotics did not interfere with the measurement of the berberine concentration. The molecular ion peaks of the complexes formed by the aptamer and berberine could be clearly observed by electrospray ionization mass spectrometry. The UV-vis absorption spectra, circular dichroism spectra, and fluorescence spectra indicated a strong interaction between berberine and the aptamer. The dissociation constant (K_d) between berberine and the aptamer was 1.91 μM. This sensor was both simple and sensitive, requiring only a 21-base oligonucleotide. It realized a visual quantitative analysis with a smartphone. This method could also be used for similar fluorescence visualization determination of aptamer-based drug molecules.