Amyloid fibrillation is associated with a great variety of human diseases, such as Alzheimer's and Huntington's diseases. A fluorescence assay for amyloid fibrillation detection and inhibitor screening was developed based on the fact that the fluorescence emission of gold nanoclusters (Au NCs) is largely enhanced upon adding amyloids, such as lysozyme amyloid fibers. A good linear relationship exists between the enhanced fluorescence intensity of Au NCs and lysozyme fiber within the concentration range of 0–0.05 mg mL⁻¹. This ultra-sensitive method can detect the protein fiber earlier than thioflavin T (THT), allowing more time for disease treatment. Furthermore, Au NCs have many advantages over the classical probe (i.e., THT), such as large Stokes shifts and low toxicity. We selected ascorbic acid as a representative inhibitor and used this method to screen inhibitors. If inhibitors are added when incubating lysozyme, the lysozyme fibrosis process will be crimped, decreasing the amount of lysozyme fibers.