Virus Neutralization Test (VNT) and Solid Phase Competitive ELISA (SPCE) are useful tools for a wide range of in vitro investigations, focused on serology studies in livestock and wild animals. After vaccination campaigns or immunogenicity studies, related to the control of Foot-and-mouth Disease (FMD), VNT and SPCE are usually performed to detect immune response of animals and, subsequently, the effectiveness of a vaccination. VNT remains the diagnostic gold standard, but various issues are related to this assay, first and foremost the manipulation of viable viruses and the related maintenance of an acceptable biosecurity level. Together with the complexity of test, mainly due to its affecting variables, this reason had often induced to find a correlation with SPCE, in order to substitute VNT with a safer and easier assay. In this study, comparing the data collected analysing sera from vaccinated animals, we evaluated the potential correlation between the two tests, focusing on the effect of homology/heterology of the strains behind the assays and those used during vaccination.