A. Dehee et al., Quantification of Epstein-Barr virus load in peripheral blood of human immunodeficiency virus-infected patients using real-time PCR, J MED VIROL, 65(3), 2001, pp. 543-552
Epstein-Barr virus (EBV) reactivation is more likely to occur in immunocomp
romised patients with subsequent higher susceptibility to EBV-associated ly
mphoproliferations. In contrast to transplant recipients, limited data are
available concerning the EBV load in HIV-infected patients, with or without
AIDS-related non-Hodgkin's lymphomas. We developed a TaqMan real-time PCR
assay, allowing both the EBV genome and a cellular gene to be quantified in
order to obtain a reliable normalized measurement of the EBV load in perip
heral blood mononuclear cells (PBMCs). With a wide 6-log(10) quantification
range and inter-assay variations of less than 24%, this quantitative PCR w
as sufficiently accurate and reproducible for routine follow-up. The EBV lo
ad was determined in PBMCs from 113 HIV-infected patients, 11 patients with
primary HIV infection and 24 HIV-seronegative healthy controls. The rates
of EBV detection were similar in the three groups. However, EBV loads were
higher in the HIV-infected group (P <0.00001) except for the patients with
primary HIV infection. Unexpectedly, EBV loads were not correlated with the
clinical stages of HIV infection or HIV replication, and did not depend on
the degree of immunodepression, as judged by CD4(+) counts. This study con
tributes towards the definition of the baseline EBV load during HIV infecti
on and stresses the broad inter-individual variability of the EBV load in H
IV-infected patients. Real-time PCR provides a useful tool that can be used
in further longitudinal studies to assess the relevance of the EBV load to
identify HIV-infected patients with a high risk of EBV-associated lymphopr
oliferations. (C) 2001 Wiley- Liss, Inc.