Au nanoparticles (NPs) possess unique physicochemical and optical properties, showing great potential in biomedical applications. Diagnostic spectroscopy utilizing varied Au NPs has become a precision tool of in vitro and in vivo diagnostic for cancer and other specific diseases. In this review, we tried to comprehensively introduce the remarkable optical properties of Au NPs, including localized surfaces plasmon resonance (LSPR), surface-enhanced Raman scattering (SERS), and metal-enhanced fluorescence (MEF). Then, we highlighted the excellent works using Au NPs for optical diagnostic applications. Ultimately, the challenges and future perspective of using Au NPs for optical diagnostic were discussed.