We report herein the development of a highly sensitive and selective colorimetric detection method for cysteine using gold nanoparticles probes. This assay relies upon the distance-dependent optical properties of gold nanoparticles, the self-assembly of cysteine on gold nanoparticles, and the interaction of a 2:1 cysteine/Cu2+ complex. In the presence of Cu2+, cysteine could rapidly induce the aggregation of gold nanoparticles, thereby resulting in red-to-blue (or purple) color change. The concentration of cysteine can be determined by monitoring with the naked eye or a UV-vis spectrometer. The present limit of detection for cysteine is 10 nM. This method exhibits excellent selectivity for cysteine over other α-amino acids, glutathione, thioglycolic acid and mercaptoethyl alcohol.