Optical characterization of a two-dimensional silver hexagram nanohole arrays was carried out. Finite-difference and time-domain method was used to calculate the transmission and localized electric field distribution of the nanohole arrays. According to the theoretical design result, focused ion beam (FIB) nanofabrication technology was employed to fabricate the hexagram nanoholes in the silver film sputter-coated on a glass substrate. The FIB fabricated nanoholes were shown to be distributed on the glass substrate regularly. Near-field scanning optical microscope (NSOM) was used to measure the electric field distribution near the surface of the nanoholes. All results show that the two-dimensional silver hexagram nanohole arrays can be fabricated by FIB successfully and that the optical intensity distribution near the nanoholes is uniform. The hexagram metallic nanoholes may be utilized for imaging, focusing and nanobiosensing.