Based on gold nanoparticles (AuNPs) and engineered DNA aptamers, we designed a novel bioassay strategy for the detection of adenosine as a small target molecule. In this design, an aptamer is engineered to consist of two pieces of random-coil like ssDNA which are respectively attached to AuNPs through their 5'-thiol-modified end. They can reassemble into the intact aptamer tertiary structure and induce nanoparticle aggregation in the presence of the specific target. Results have demonstrated that gold nanoparticles can effectively differentiate these two different DNA structures via their characteristic surface plasmon resonance-based color change. With this method, adenosine can be selectively detected in the low micromolar range, which means that the strategy reported here can be applicable to the detection of several other small target molecules.