Rational design of detection strategy is the key to high-performance fluorescence analysis. In this article, we found that the glucose-induced limitations can greatly enhance the fluorescence of functionalized carbon nanoparticles (CNPs), which are synthesized through one-step thermal pyrolysis method using phenylboronic acid derivative as the precursors. The glucose can assembly onto the surface of the CNPs to form a "shell", limiting the surfaces' intramolecular rotation and reducing non-radiative decay, which hence resulted in enhanced fluorescence of the CNPs. Under optimal conditions, the fluorescence intensity of the CNPs is nearly 70-fold enhanced, and the method has low detection limit (10μM) and linear response in the concentration range from 50μM to 2000μM. Based on this interesting "target-triggered limitation-induced fluorescence enhancement" phenomenon, a simple and effective non-enzymatic fluorescence enhancement method was developed and successfully applied to the determination of glucose in spiked serum samples. This work provides new insight into the design of fluorescence-enhanced detection strategies based on the limitation-induced property.