Polytetrafluoroethylene (PTFE) is a fascinating electret material widely used for energy harvesting and sensing, and an enhancement in the performance could be expected by reducing its size into nanoscale because of a higher surface charge density attained. Hence, the present study demonstrates the use of nanofibrous PTFE for high-performance self-powered wearable sensors. The nanofibrous PTFE is fabricated by electrospinning with a suspension of PTFE particles in dilute polyethylene oxide (PEO) aqueous solution, followed by a thermal treatment at 350 °C to remove the PEO component from the electrospun PTFE-PEO nanofibers. The obtained PTFE nanofibrous membrane exhibits good air permeability with pressure drop comparable to face masks, excellent mechanical property with tensile strength of 3.8 MPa, and stable surface potential of − 270 V. By simply sandwiching the PTFE nanofibrous membrane into two pieces of conducting carbon clothes, a breathable, flexible, and high-performance nanogenerator (NG) device with a peak power of 56.25 μW is constructed. Remarkably, this NG device can be directly used as a wearable self-powered sensor for detecting body motion and physiological signals. Small elbow joint bending of 30°, the rhythm of respiration, and typical cardiac cycle are clearly recorded by the output waveform of the NG device. This study demonstrates the use of electrospun PTFE nanofibrous membrane for the construction of high-performance self-powered wearable sensors.