A convenient semi-quantitative method for the diagnosis of Epstein-Barr virus reactivation

A semi-quantitative determination of Epstein-Barr virus (EBV) viremia has been devised. Peripheral blood mononuclear cells are recovered by Ficoll gradient and numerated. Five microl aliquots of recovered cell suspension and 5 microl of two standard dilutions (containing 500 and 100 cells, respectively) are subjected to a nested polymerase chain reaction (PCR). This technique has been evaluated over 3 years for the follow-up of 45 patients attending the Bone Marrow Transplantation Unit of the "Centre Hospitalier et Universitaire de Nancy". EBV reactivation was diagnosed in 13 patients (28%). Positivity of PCR for 100 cells was found in 9 patients of whom 6 developed lymphoma or lymphoproliferative disorder. This technique is easy to perform and doesn't necessitate any specific material besides the one necessary for routine genic amplification.