Effective biosensor devices for detection of glucose based on carbohydrate polymer nanohydrogels have been scarcely reported to date. In an attempt to construct a more functional device for detection of blood glucose, CdTe quantumdots (QDs) and glucose oxidase (GOx) were immobilizedwithin the well functionalized superabsorbent nanohydrogel prepared from the nanoparticles (b50 nm) of gum tragacanth (GT) polysaccharide. The biosensor was able to perform enzyme-catalyzed oxidation of glucose to produce H2O2, a product that subsequently quenched the fluorescence emission. The quenching rate depends on the glucose concentration, the detection limit was 0.5 mM, and the Michaelis–Menten constant (KM) was an excellent value of 2.2 mM. The prepared biosensor was used for the enzymatic detection of blood glucose concentration in the real serum samples which exhibited satisfactory reproducibility and accuracy. Therefore, the high immobilization for QDs and GOx in this superabsorbent nanohydrogelmatrix can be a suitable diagnostic device for biomedical applications.