[Display omitted] • A phosphorescence sensing system for alkaline phosphatase was developed. • It is based on the interaction between Mn:ZnS quantum dots and pyrophosphate. • The sensing system is low-cost, non-toxic and eco-friendly. • A phosphorescence-based "INHIBIT" logic gate for alkaline phosphatase was presented. This paper presents a highly sensitive sensing system for alkaline phosphatase by room temperature phosphorescence of Mn doped ZnS quantum dots and pyrophosphate. The sensing system has intense room temperature phosphorescence emission in the absence of alkaline phosphatase. The phosphorescence is quenched gradually with the addition of alkaline phosphatase. The emission "on" without alkaline phosphatase may be attributed to the increased probability of charge transfer from one of surface traps to the dopant bands of another resulted from the shortened dot-to-dot distance by the strong chelation of pyrophosphate and Zn2+ ion and the hydrogen bonding between pyrophosphate and β-cyclodextrin. The addition of alkaline phosphatase causes pyrophosphate hydrolyzed to orthophosphate and the dot-to-dot distance of quantum dots back to the normal, and then the phosphorescence "off". The factors affecting the sensing system performance were also optimized. Under the optimal experimental conditions, the linear range for alkaline phosphatase is determined as 0.2−10 U/L with a LOD at 0.045 U/L. The recovery of human serum was determined from 93.75%–103.03%, indicating a potential application in biomedical diagnosis. Furthermore, an RTP-based "INHIBIT" logic gate using the doped ZnS quantum dots was also presented. [ABSTRACT FROM AUTHOR]