For the first time, boronic acid–modified polymer dots (B-PDs) were fabricated by a “synthesis-modification integration” route using polyethylenimine (PEI) and phenylboronic acid as precursors. Under optimized preparation conditions, the B-PDs exhibited an average size of 2.2 nm, good water solubility, and high fluorescence quantum yield of 8.69%. The B-PDs showed reversible fluorescence response in acid solutions (blue emissions) and alkaline solutions (green emissions). The fluorescence emissions of B-PDs demonstrated an obvious red shift with varying the pH value from 1 - 13. Moreover, glucose could assemble on the surface of B-PDs due to the reversible reaction between boronic acid and cis -diols, which resulted in a blue shift of emission wavelength and an obvious increase of FL intensity at λ ex  = 380 nm based on the aggregation-induced enhancement effect. The glucose sensing method was thus developed in the range 0.0001 - 1.0 mol L −1 . Applications to real human blood and glucose injection samples demonstrated satisfactory results. The B-PDs based on the analytical method display good selectivity, wide detection range, and simplicity in preparation and detection, implying promising applications as a practical platform for biosensing. Graphical abstract