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Cellular normalization of viral DNA loads on whole blood improves the clinical management of cytomegalovirus or Epstein Barr virus infections in the setting of pre‐emptive therapy
Authors:Céline Bressollette‐Bodin  Marianne Coste‐Burel  Bernard Besse  Elisabeth André‐Garnier  Virginie Ferre  Berthe‐Marie Imbert‐Marcille
Affiliation:1. Université de Nantes, Nantes Atlantique Universités, EA4271 ? Immunovirology and Genetic Polymorphisms ?, IFR26, Nantes, France;2. Virology Laboratory, Institut de Biologie du Centre Hospitalier Universitaire de Nantes, Nantes, France
Abstract:Two quantitative duplex real‐time PCR assays were developed for co‐amplification of human albumin and cytomegalovirus (CMV) or Epstein Barr virus (EBV) genes after automated extraction on whole blood, and compared two units for expressing viral DNA loads (copies per ml of blood or per 106 peripheral blood leukocytes (PBLs)) on 1,138 positive samples. Both PCRs were characterized by high sensitivity, reproducibility, and linear range. Automated extraction by a MagNA Pure LC Instrument was shown to be more efficient when peripheral blood cell count was inferior to 5 × 109 PBLs/L. Albumin co‐amplification allows the detection of PCR inhibitors and normalization of viral load according to the number of cells calculated in the sample. The two ways of expressing viral load results were highly correlated, but quantitative differences varied in relation to variations of blood cell count. As these two viruses are highly cell associated, viral loads can be underestimated in patients with leucopenia. In the setting of pre‐emptive strategies during CMV infection, the units in which results are expressed can influence clinical management, as illustrated in this article. J. Med. Virol. 81:90–98, 2009. © 2008 Wiley‐Liss, Inc.
Keywords:cytomegalovirus  Epstein Barr virus  real‐time PCR  viral loads
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