Based on inducing the aggregation of gold nanoparticles (AuNPs), a simple colorimetric method with high sensitivity and selectivity was developed for the sensing of thiocyanate (SCN(-)) in aqueous solutions. Citrate-capped AuNPs were prepared following a classic method and Tween 20 was subsequently added as a stabilizer. With the addition of SCN(-), citrate ions on AuNPs surfaces were replaced due to the high affinity between SCN(-) and Au. As a result, Tween 20 molecules adsorbed on the AuNPs surfaces were separated and the AuNPs aggregated. The process was accompanied by a visible color change from red to blue within 5 min. The sensing of SCN(-) can therefore be easily achieved by a UV-vis spectrophotometer or even by the naked eye. The potential effects of relevant experimental conditions, including concentration of Tween 20, pH, incubation temperature and time, were evaluated to optimize the method. Under optimized conditions, this method yields excellent sensitivity (LOD = 0.2 μM or 11.6 ppb) and selectivity toward SCN(-). Our attempt may provide a cost-effective, rapid and simple solution to the inspection of SCN(-) ions in saliva and environmental aqueous samples.