title: 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.
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 10(6) 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 x 10(9) 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.
authors.affiliation: Université de Nantes, Nantes Atlantique Universités, EA4271 << Immunovirology and Genetic Polymorphisms >>, IFR26, Nantes, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Bressollette-Bodin
authors.forename: Marianne
authors.initials: M
authors.lastname: Coste-Burel
authors.forename: Bernard
authors.initials: B
authors.lastname: Besse
authors.forename: Elisabeth
authors.initials: E
authors.lastname: André-Garnier
authors.forename: Virginie
authors.initials: V
authors.lastname: Ferre
authors.forename: Berthe-Marie
authors.initials: BM
authors.lastname: Imbert-Marcille
mesh_terms: D019090:Case Management; D003587:Cytomegalovirus / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000628:therapy*; D004279:DNA, Viral / Q000097:blood*; D020031:Epstein-Barr Virus Infections / Q000628:therapy*; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D004279:DNA, Viral