Fluorometric aptamer based determination of adenosine triphosphate based on deoxyribonuclease I-aided target recycling and signal amplification using graphene oxide as a quencher

Abstract

The authors describe an aptamer based fluorometric assay for the determination of ATP. It is based on deoxyribonuclease I-aided target recycling and signal amplification. The DNA probe consists of two regions (sequences) that represent the capture probe and the signal probe, respectively. In the absence of ATP, the probe is adsorbed by the surface of graphene oxide (GO) via π-stacking interactions. This results in quenching of the fluorescent label (carboxyfluorescein) and protects it from being cleaved by DNase. Upon adding ATP, the probe will be repelled by GO because ATP binds to the aptamer. This triggers an increase in fluorescence as measured at excitation/emission wavelengths of 480/514 nm. The detection limit is as low as 0.2 nM, and the calibration plot is linear in the 10 to 400 nM ATP concentration range. The assay is specific and sensitive, and in our perception has a large potential in terms of detecting other species including pathogenic microorganisms, small molecules, metal ions, and proteins.