A large-area efficient surface enhanced Raman scattering (SERS) substrate was deposited by Langmuir-Blodgett (LB) assembly and followed by sputter coating process. The interparticle distance of silica nanospheres was convenient and readily regulated by controlling the surface pressure. After sputter coating with a small amount of gold, the nanocomposite film was formed. The surfaces of nanostructures were characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The applicability of the nanostructures as SERS substrates was investigated by adsorption of crystal violet (CV) molecules. Different SERS effects were observed on deposited gold nanostructure upon large-area silica nanospheres monolayer with diameter of 250,570 and 800 nm. It was also discussed the nanospheres with diameter of 250 nm for the fabrication of ordered arrangement of monolayers at surface pressures of 10-30 mN/m. These substrates with the close-packed nanospheres and sputtering of gold were found to exhibit high and uniform enhancement of the Raman signal across the entire surface. The averaged surface enhancement factor (ASEF) was calculated and discussed. The fabricated nanocomposite structures could be utilized as low-cost SERS-active substrates for biomedical and analytical chemistry field. (C) 2014 Elsevier B.V. All rights reserved.