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. J. Med. Virol. 86:851–856, 2014 . © 2013 Wiley Periodicals, Inc.