We synthesized a fluorescent sensor for detection of pyrophosphate (PPi) and alkaline phosphatase (ALP) based on copper/silver nanoclusters (DNA-Cu/Ag NCs) with single-stranded DNA as template. The whole synthesis process was only 30 min. When PPi was added to the reaction liquid for the preparation of DNA-Cu/Ag NCs, the fluorescence of DNA-Cu/Ag NCs would be quenched due to the strong complexation between PPi and Cu 2+. As ALP could catalyze the hydrolysis of PPi to Pi, the complexation between Cu 2+ and PPi would be destroyed and the fluorescence intensity would increase. Based on this principle, we developed a new method for detecting PPi and ALP with a detection limit of 136.7 nmol/L and 0.01 U/mL, respectively. Finally, by detecting the PPi in human serum, the diagnostic ability and practical application of the method were verified.
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