In this work, a straightforward one-step hydrothermal method was efficiently applied to prepare boronic acid-functionalized N-doped carbon dots (BN-CDs) using 3-aminobenzeneboronic acid and ascorbic acid as the starting materials. The BN-CDs exhibited a size distribution ranging from 1.5 nm to 4.0 nm, demonstrated good fluorescence stability and a high fluorescence quantum yield of 26.03 %. Glucose could accumulate on the surface of BN-CDs by covalently binding boric acid groups to glucose cis-glycol, thereby leading to the selective fluorescence quenching performance. A sensitive sensing platform for glucose detection was established in the linear range of 8-940 μmol/L with the detection limit (LOD) of 2.4 μmol/L. Satisfactory results were obtained in three kinds of sample analysis. It is worth mentioning that the incubation time of the proposed sensing platform is much shorter than that of most reported elsewhere. The BN-CDs based fluorescence detection method demonstrates good feasibility in practical applications.