A new and efficient nitric oxide (NO) detection method based on the DAF-FM which is a kind of NO probes was proposed. The incorporation of gold nanoparticles significantly increased the fluorescence intensity of the fluorescent probe after reacting with NO. The new fluorescent probes composed of DAF-FM and gold nanoparticles have dozens of times higher fluorescence intensity than that without gold nanoparticles does. The sensitivity of NO detection has been significantly improved. The new fluorescent probe can be reused for more than 24 times, overcoming the single-use problem of DAF-FM and achieving the sustainable reuse of commercial nitric oxide detection.