We report a simple method to efficiently improve the detection limit of surface plasmon resonance in periodic metallic nanostructures by using small angle illumination and spectral integration analysis. The large-area gold nanoslit arrays were fabricated by thermal-annealing template-stripping method with a slit width of 60 nm and period of 500 nm. The small angle illumination induced a resonant coupling between surface plasmon mode and substrate mode. It increased ~2.24 times intensity sensitivity at 5.5° incident angle. The small-angle illumination also resulted in multiple resonant peaks. The spectral integration method integrated all changes near the resonant peaks and increased the signal to noise ratio about 5 times as compared to single-wavelength intensity analysis. Combining both small angle and spectral integration, the detection limit was increased to one order of magnitude. The improvement of the detection limit for antigen-antibody interactions was demonstrated.