Abstract: Methods for the rapid detection of the presence or absence of Epstein Barr Virus (EBV) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers and probes targeting EBV, and kits are provided that are designed for the detection of target regions of EBV. Also described are kits, reaction mixtures, and oligonucleotides (e.g., primer and probe) for the amplification and detection of EBV. Also described are primers and probes that detect different regions of EBV, and can be employed in a dual target assay for simultaneously detecting two different and non-overlapping target regions of EBV.