Nano-porous gold membrane was fabricated by chemical etching method in the present work. The structures and the unique optical properties of the membrane were studied by scanning electron microscope and spectrometer. We creatively used the nano-porous gold membrane as the sensing layer for surface plasmon resonance biosensor. The results showed that, compared to the traditional evaporated gold film, the nano-porous gold membrane has a unique effect of localized surface plasmon resonance. The sensitivity of detecting the biological agents was obviously increased by the surface plasmon resonance biosensor assembling the nano-porous gold membrane. The fabrication method of the nano-porous gold membrane is simple and low-cost, so it absolutely can replace the use of traditional evaporated gold film.Sensitivity