Arch Virol (2000) 145: 2211–2216
A convenient semi-quantitative method for the diagnosis
of Epstein-Barr virus reactivation
, A. S. Carret
, N. Pascal
, B. Rihn
, P. Bordigoni
, and A. Le Faou
Unité Mixte de Recherche, 7565 UHP-CNRS, Laboratoire de Bactériologie-Virologie,
Faculté de Médecine, Vandœuvre-lès-Nancy, France
Unité de Transplantation Médullaire, Hôpital d’Enfants, CHU Nancy-Brabois,
Accepted May 2, 2000
Summary. 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 l aliquots of recovered cell suspension and
5 l 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.
Epstein-Barr Virus (EBV) is associated with 100% of lymphomas occuring in
bone marrow grafted patients [1, 7, 8, 16]. This deadly disease is observed mainly
in patients receiving marrow from an unrelated donor, due to the necessity of
an acute immunodepression to overcome graft-versus-host disease [3, 6, 13, 16].
Such a malignant evolution occurs within the 6 months following transplantation.
Therefore, a follow-up of these patients during this period is necessary in order
to ensure early diagnosis and treatment of EBV related lymphoma.
In immunocompromised patients, information obtained from serology is lim-
ited  and for detection of reactivation, direct diagnosis is recommended .
Gene ampliﬁcation is the only method that combines sensitivity, and rapidity and