The fabrication of two-dimensional (2D) metal-organic frameworks (MOFs) and Prussian blue analogues (PBAs) combines the advantages of 2D materials, MOFs and PBAs, resolving the poor electronic conductivity and slow diffusion of MOF materials for electrochemical applications. In this work, 2D leaflike zeolitic imidazolate frameworks (Co-ZIF and Fe-ZIF) as sacrificial templates are in situ converted into PBAs, realizing the successful fabrication of PBA/ZIF nanocomposites on nickel foam (NF), namely, CoCo-PBA/Co-ZIF/NF, FeFe-PBA/Fe-ZIF/NF, CoFe-PBA/Co-ZIF/NF, and Fe/CoCo-PBA/Co-ZIF/NF. Such fabrication can effectively reduce transfer resistance and greatly enhance electron- and mass-transfer efficiency due to the electrochemically active PBA particles and NF substrate. These fabricated electrodes as multifunctional sensors achieve highly selective and sensitive glucose and HO biosensing with a very wide detective linear range, extremely low limit of detection (LOD), and good stability. Among them, CoFe-PBA/Co-ZIF/NF exhibits the best sensing performance with a very wide linear range from 1.4 μM to 1.5 mM, a high sensitivity of 5270 μA mM cm, a low LOD of 0.02 μM (/ = 3), and remarkable stability and selectivity toward glucose. What is more, it can realize excellent detection of glucose in human serum, demonstrating its practical applications. Furthermore, this material as a multifunctional electrochemical sensor also manifests superior detection performance against hydrogen peroxide with a wide linear range of 0.2-6.0 mM, a high sensitivity of 196 μA mM cm, and a low limit of detection of 1.08 nM (/ = 3). The sensing mechanism for enhanced performance for glucose and HO is discussed and proved by experiments in detail.