Poly(vinylidene fluoride) (PVDF) electrospun nanofibrous membranes (NFMs) have good piezoelectric properties compared to other polymer materials due to their electroactive phase. However, there is a limit to the content of the electroactive phase (FEA) in PVDF, which restricts the further improvement of PVDF NFMs' piezoelectric properties. This research prepared PVDF multilayer nanofibrous membranes (MNFMs) with a novel 3D structure by assembling electrospun PVDF nanofibers and electrospraying PVDF microspheres. The various properties of PVDF microspheres and PVDF MNFMs, including their crystalline structure, morphology, and piezoelectric properties, were discussed in detail. The addition of PVDF microspheres to PVDF NFMs hardly changed the FEA in PVDF MNFMs. However, the piezoelectric properties of PVDF MNFMs significantly increased compared to those of PVDF NFMs. PVDF MNFMs demonstrate good piezoelectric sensitivity and can detect the stress force as small as 0.25 N and finger tapping. The maximum output voltage of PVDF MNFMs was 112 mV, achieved when applying a force of 0.25 N at a frequency of 110 Hz. This value is three times higher than that of PVDF NFMs. [ABSTRACT FROM AUTHOR]