The aim was to evaluate two commercial enzyme immunoassay (EIA) Lyme Borreliosis IgG kits namely the NovaLisa Borrelia burgdorferi IgG ELISA (recombinant) and the Euroimmun Anti-Borrelia (whole cell lysate) plus VlsE ELISA IgG compared to the screening kit currently utilised namely the MarDx Diagnostics (Trinity Biotech) Borrelia burgdorferi EIA IgG and IgM whole cell lysate and an in-house western immunoblot IgG using whole cell lysates of B.burgdorferi and B.afzelii. 267 previously tested and stored serum samples were tested in the EIAs and 226 of these were also tested in western immunoblots. These specimens also included a specificity panel (leptospirosis, syphilis, H. pylori, EBV, ANA and rheumatoid factor positives), four dilution series and one sample repeated ten times. The recombinant antigen NovaLisa IgG had an overall agreement of only 60% with the whole cell antigen total antibody MarDx while the whole cell plus VlsE Euroimmun IgG showed a higher agreement of 72%. This was expected as the comparison with western immunoblot using the CDC criteria for positive IgG blots of five specific bands gave MarDx only a 45% agreement, NovaLisa 88% agreement and Euroimmun 69% agreement. The dilution series indicated that Eurimmun IgG was slightly more sensitive (one doubling dilution) for two specimens and NovaLisa IgG more sensitive for one specimen so they could not be separated by this method. The specificity panel showed NovaLisa IgG was the most specific at 96% with MarDx 88% and Euroimmun 83%. Repeatability gave coefficients of variation of 5% for Euroimmun and 15% for NovaLisa but this may have indicated operator variation at the time of testing. With a greater specificity compared to western immunoblot and known cross reactive specimens, the NovaLisa IgG recombinant EIA was considered the most accurate kit to use as a screening test for Lyme borreliosis.