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Surface-enhanced Raman spectroscopy (SERS) became a useful analytical technique with the development of appropriate metallic substrates. The need for SERS substrates that immobilize metallic nanoparticles prompted this work to search for an appropriate material. This work presents the preparation, characterization and application of a SERS substrate for crystal violet (CV) detection, as the probe molecule. The inner layer of the substrate is a thin film of the fungal β-D-glucan, botryosphaeran, covered by a thin layer of silver nanopartic