A simple and robust 2-dimensional cowpea mosaic virus particle (CPMV) sensor was fabricated using a nanoparticle-imprinted polymer film. Well-known for its biocompatibility and controllable synthesis, polypyrrole (PPyr) was chosen as the main polymer matrix for the sensor. Prior to the anodic polymerization via cyclic voltammetry (CV) and cross-linking of pyrrole, the formation of a stable monomer-virus template complex due to hydrogen-bonding interactions was significant in ensuring that both components will be bound to the surface and complementary virus-sized cavities will be formed after subsequent extraction of CPMV. The stable formation of PPyr-CPMV films was characterized using atomic force microscopy (AFM), contact angle measurements, and X-ray photoelectron spectroscopy (XPS). Furthermore, an antibody-based dot blot study was used to visually verify the presence of the virus nanoparticles in the polymer film. Through a combination of acid treatments and electrochemical elution steps, the virus nanoparticles were extracted from the composite film thus revealing cavities with morphological features and functional groups complementary to the virus template as characterized by electrochemical impedance spectroscopy (EIS). The same technique was then used to measure changes in the charge transfer resistance and impedance of the films after virus extraction and subsequent rebinding of the virus nanoparticles. In order to analyze the selectivity of the electrochemical sensor, the imprinted film was tested against the Qβ virus, a similarly structured virus nanoparticle. The resulting nanoparticle imprinted sensor will find significant potential applications in monitoring viruses in agricultural settings and even in investigating the interactions of various surface-modified virus nanoparticles.