The Enzygnost Anti-EBV/IgG shows good performance for populations not defined by age. Samples from 349 pediatric outpatients were screened with the assay and with immunofluorescence assay (IFA) as the gold standard. The Enzygnost assay showed a sensitivity of 85%. Ten and one-tenth percent of EBV seropositive samples were classified as intermediate, and 4.9% were falsely determined as seronegative when compared to the standard IgG anti-virus capsid antigen (VCA) IFA. Our data suggest a limited sensitivity of the assay with samples from pediatric patients.