Comparison of EBV DNA viral load in whole blood, plasma, B-cells and B-cell culture supernatant

J Med Virol. 2014 May;86(5):851-6. doi: 10.1002/jmv.23858. Epub 2013 Nov 22.

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.

Keywords: B-cells culture supernatant; EBV DNA load; blood specimens.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / virology*
  • Blood / virology*
  • Cell Culture Techniques
  • Clinical Laboratory Techniques / methods*
  • Culture Media
  • DNA, Viral / isolation & purification*
  • Drug Monitoring / methods
  • Epstein-Barr Virus Infections / diagnosis*
  • Epstein-Barr Virus Infections / virology*
  • HIV Infections / complications
  • Humans
  • Lymphoma, B-Cell / virology
  • Real-Time Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Viral Load*
  • Virology / methods

Substances

  • Culture Media
  • DNA, Viral