Abstract: Disclosed are compositions, methods, and kits that can be used to Epstein-Ban virus (EBV) in a sample undergoing testing. Nucleic acids of EBV can be isolated, amplified and detected with specificity by real-time PCR, and without interference from non-EBV organisms. In some embodiments, nucleic acids used for amplification are isolated from human blood, or blood products. Nucleic acid isolation, amplification and detection steps can all be carried out using an automated instrument.