Serological tests detecting antibodies for Epstein-Barr virus (EBV) antigens are frequently used to define infection status. Several new automated assays are available for this purpose. We compared the performance of Architect, Immulite, Vidas, and Euroimmune immunofluorescence assays (IFA)/enzyme-linked immunosorbent assays (ELISA) for the detection of EBV viral capsid antigen (VCA) immunoglobulin M (IgM), VCA IgG, Epstein-Barr nuclear antigen (EBNA)-1 IgG. The routine diagnosis of EBV in our laboratory is done by anti-EBV VCA IgM IFT, anti-EBV VCA IgG IFT, and anti-EBNA-1 IgG ELISA (Euroimmune) Kits. Samples were tested with EBV Kits of Architect, Immulite, and Vidas for anti-VCA IgM, anti-VCA IgG, and anti-EBNA-1 IgG. The agreement between assays was calculated for each marker individually and for the determination of the EBV infection profile, based on the combination of three markers. BIOCHIP Sequence EBV (Avidity test) and/or EUROLINE EBV Profile 2 (IgG/IgM) were used as confirmatory assays to resolve discrepancies. The best concordance for VCA IgM detection was between Immulite and Vidas; for VCA IgG and EBNA-1 IgG were between Architect andVidas. The sensitivities and specificities for VCA IgM were 97% and 88% for IFA, 100% and 94% for Architect, 100% and 99% for Vidas, and 100% and 100% for Immulite, respectively. The most problematic marker was EBNA-1 IgG with a 68.1% specificity by Immulite. Vidas panel had a perfect performance (100%) for determining all EBV profiles. Overall, evaluated assays had comparable performance. There were more discordant VCA IgG and EBNA-1 IgG results than VCA IgM results.The agreement between Architect and Vidas was better than other assays.