Receptor-functionalized membranes provide a new paradigm for developing mechanical biosensors. However, the lower sensitivity of these biosensors is still a challenge. In this research, a sensitive and thin (1.5 [Formula: see text]m) gold nanoparticles-polydimethylsiloxane (AuNPs-PDMS) membrane is fabricated by reducing HAuCl4. Experimental results reveal that surface stress deforms the AuNPs-PDMS membrane, further increasing the resistance due to the tunneling mechanism and conductive percolation. During the functional process, the resistance change is consistent with theoretical prediction. The relative resistance change demonstrates a linear relationship with the glucose concentration. The biosensors exhibit a wider linear range for glucose from 0 to 30[Formula: see text]mM with a lower detection limit of 0.035[Formula: see text]mM. AuNPs-PDMS thin membrane-based surface stress biosensor shows promising application prospects to detect biomolecules.