title: Comparison of EBV DNA viral load in whole blood, plasma, B-cells and B-cell culture supernatant.
abstract: Epstein-Barr virus (EBV) genome quantitation in whole blood is used widely for therapeutic monitoring of EBV-associated disorders in immunosuppressed individuals and in patients with EBV-associated lymphoma. However, the most appropriate biological material to be used for EBV DNA quantitation remains a subject of debate. This study compare the detection rate and levels of EBV DNA from whole blood, plasma, enriched B-cells, and B-cell short-term culture supernatant using quantitative real-time PCR. Samples were collected from 33 subjects with either HIV infection or B-cell lymphoma. Overall, EBV DNA was detected in 100% of enriched B-cell samples, in 82% of B-cell culture supernatants, in 57% of plasma, and 42% of whole blood samples. A significant correlation for EBV viral load was found between enriched B-cell and B-cell culture supernatant material (ρ = 0.92; P < 0.0001), but no significant correlation existed between EBV DNA levels in whole blood and enriched B-cells (ρ = -0.02; P = 0.89), whole blood and plasma (ρ = 0.24; P = 0.24), or enriched B-cells and plasma (ρ = 0.08; P = 0.77). Testing of enriched B-cells appeared to be the most sensitive method for detection of EBV DNA as well as for exploration of the cellular reservoir. Quantitation of EBV DNA in plasma and B-cell culture supernatant may be of interest to assess EBV reactivation dynamics and response to treatment as well as to decipher EBV host-pathogen interactions in various clinical scenarios.
authors.affiliation: INSERM U1058, University of Montpellier 1, Montpellier, France; Montpellier Hospital Centre, Institute of Biotherapies Research, Laboratory of Human Rare Circulating Cells, Montpellier, France; Montpellier Hospital Centre, Department of Bacteriology and Virology, Paris, France.
authors.forename: David Eric
authors.initials: DE
authors.lastname: Ouedraogo
authors.forename: Karine
authors.initials: K
authors.lastname: Bollore
authors.forename: Johannes
authors.initials: J
authors.lastname: Viljoen
authors.forename: Vincent
authors.initials: V
authors.lastname: Foulongne
authors.forename: Jacques
authors.initials: J
authors.lastname: Reynes
authors.forename: Guillaume
authors.initials: G
authors.lastname: Cartron
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Vendrell
authors.forename: Philippe
authors.initials: P
authors.lastname: Van de Perre
authors.forename: Edouard
authors.initials: E
authors.lastname: Tuaillon
mesh_terms: D001402:B-Lymphocytes / Q000821:virology*; D001769:Blood / Q000821:virology*; D018929:Cell Culture Techniques; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003470:Culture Media; D004279:DNA, Viral / Q000302:isolation & purification*; D016903:Drug Monitoring / Q000379:methods; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis* / Q000821:virology*; D015658:HIV Infections / Q000150:complications; D006801:Humans; D016393:Lymphoma, B-Cell / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019562:Viral Load; D014773:Virology / Q000379:methods
chemical_list: D003470:Culture Media; D004279:DNA, Viral
keywords: B-cells culture supernatant; EBV DNA load; blood specimens