Indian Pediatrics

, Volume 52, Issue 8, pp 681–685 | Cite as

Plasma Epstein-Barr virus (EBV) DNA as a biomarker for EBV-associated Hodgkin lymphoma

  • Veronique Dinand
  • Anupam Sachdeva
  • Sanghamitra Datta
  • Sunita Bhalla
  • Manas Kalra
  • Chand Wattal
  • Nita Radhakrishnan
Research Paper



To assess plasma Epstein-Barr virus (EBV) DNA as a biomarker of tumour burden at diagnosis and during therapy in children with Hodgkin lymphoma.


Case-control study, with prospective follow-up of the Hodgkin lymphoma cohort (2007-2012).


Pediatric Hematology Oncology unit of a tertiary care hospital in Delhi.


Thirty children with Hodgkin lymphoma and 70 sex and age-matched controls (benign lymphadenopathy 19, non-lymphoid malignancy 29, Burkitt lymphoma 5, healthy children 17).


Positive EBV-staining on immunohistochemistry was defined as EBV-associated Hodgkin lymphoma. Plasma EBV real-time quantitative polymerase chain reaction (PCR) was tested at presentation, after first and last chemotherapy cycles, and on follow-up.

Main outcome measures

Plasma EBV quantitative PCR was compared between cases and controls. Its kinetics was assessed during and after chemotherapy.


Results: EBV quantitative PCR was positive in 19 (63%) Hodgkin lymphoma cases (range 500–430,000 copies/mL), with 87.5% accuracy (kappa=0.69) as compared with EBVimmunohistochemistry. Sensitivity and specificity of the quantitative PCR were 87.5% and 81.8%, respectively. Only boys showed positive EBV immunohistochemistry and/or quantitative- PCR positivity. All controls were quantitative-PCR negative. All quantitative-PCR positive cases with follow-up blood sample showed EBV clearance after the first cycle. A quantitative-PCR negative case in long-term remission became positive at relapse. EBV status did not influence survival.


Plasma EBV-DNA, detectable in EBV-associated Hodgkin lymphoma, becomes undetectable early after initiating therapy. It can be used as a biomarker of treatment response in EBV-associated Hodgkin lymphoma.


Follow-up Immuno-histochemistry Outcome 


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  1. 1.
    Glaser SL, Lin RJ, Stewart SL, Ambinder RF, Jarrett RF, Brousset P, et al. Epstein-Barr virus-associated Hodgkin’s disease: epidemiologic characteristics in international data. Int J Cancer. 1997;(70):375–82.CrossRefPubMedGoogle Scholar
  2. 2.
    Dinand V, Dawar R, Arya LS, Unni R, Mohanty B, Singh R. Hodgkin’s lymphoma in Indian children: prevalence and significance of Epstein-Barr virus detection in Hodgkin’s and Reed Sternberg cells. Eur J Cancer. 2007;(43):161–8.CrossRefPubMedGoogle Scholar
  3. 3.
    Gallagher A, Armstrong AA, MacKenzie J, Shield L, Khan G, Lake A, et al. Detection of Epstein-Barr virus (EBV) genomes in the serum of patients with EBV-associated Hodgkin’s disease. Int J Cancer. 1999;(84):442–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Gandhi MK, Lambley E, Burrows J, Dua U, Elliott S, Shaw PJ, et al. Plasma Epstein-Barr virus (EBV) DNA is a biomarker for EBV-positive Hodgkin’s lymphoma. Clin Cancer Res. 2006;(12):460–4.CrossRefPubMedGoogle Scholar
  5. 5.
    Wagner HJ, Schlager F, Claviez A, Bucsky P. Detection of Epstein-Barr virus DNA in peripheral blood of paediatric patients with Hodgkin’s disease by real-time polymerase chain reaction. Eur J Cancer. 2001;(37):1853–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Lister TA, Crowther D, Sutcliffe SB, Glatstein E, Canellos GP, Young RC, et al. Report of a committee convened to discuss the evaluation and staging of patients with Hodgkin’s disease: Cotswolds meeting. J Clin Oncol. 1989;(7):1630–6.PubMedGoogle Scholar
  7. 7.
    Harris NL. Hodgkin’s disease: classification and differential diagnosis. Mod Pathol. 1999;(12):159–75.PubMedGoogle Scholar
  8. 8.
    Naresh KN, Johnson J, Srinivas V, Soman CS, Saikia T, Advani SH, et al. Epstein-Barr virus association in classical Hodgkin’s disease provides survival advantage to patients and correlates with higher expression of proliferation markers in Reed-Sternberg cells. Ann Oncol. 2000;(11):91–6.CrossRefPubMedGoogle Scholar
  9. 9.
    Karnik S, Srinivasan B, Nair S. Hodgkin’s lymphoma: immunohistochemical features and its association with EBV LMP-1. Experience from a South Indian hospital. Pathology. 2003;(35):207–11.PubMedGoogle Scholar
  10. 10.
    Sinha M, Rao CR, Shafiulla M, Appaji L, Bs AK, Sumati BG, et al. Cell-free Epstein-Barr viral loads in childhood Hodgkin lymphoma: a study from South India. Pediatr Hematol Oncol. 2013;(30):537–3.CrossRefPubMedGoogle Scholar
  11. 11.
    Jarrett RF. Viruses and lymphoma/leukaemia. J Pathol. 2006;(208):176–86.CrossRefPubMedGoogle Scholar
  12. 12.
    Razzouk BI, Srinivas S, Sample CE, Singh V, Sixbey JWl. Epstein-Barr virus DNA recombination and loss in sporadic Burkitt’s lymphoma. J Infect Dis. 1996;(173):529–35.CrossRefPubMedGoogle Scholar
  13. 13.
    Gan YJ, Razzouk BI, Su T, Sixbey JW. A defective, rearranged Epstein-Barr virus genome in EBER-negative and EBER-positive Hodgkin’s disease. Am J Pathol. 2002;(160):781–6.PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Kanakry JA, Li H, Gellert LL, Lemas MV, Hsieh WS, Hong F, et al. Plasma Epstein-Barr virus DNA predicts outcome in advanced Hodgkin lymphoma: correlative analysis from a large North American cooperative group trial. Blood. 2013;(121):3547–53.PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Keegan TH, Glaser SL, Clarke CA, Gulley ML, Craig FE, Digiuseppe JA, et al. Epstein-Barr virus as a marker of survival after Hodgkin’s lymphoma: a population-based study. J Clin Oncol. 2005;(23):7604–13.CrossRefPubMedGoogle Scholar
  16. 16.
    Jarett RF, Stark GL, White J, Angus B, Alexander FE, Krajewski AS, et al. Scotland and Newcastle Epidemiology of Hodgkin Disease Study Group. Impact of tumor Epstein-Barr virus status on presenting features and outcome in age-defined subgroups of patients with classic Hodgkin lymphoma: a population-based study. Blood. 2005;(106):2444–51.CrossRefGoogle Scholar

Copyright information

© Indian Academy of Pediatrics 2015

Authors and Affiliations

  • Veronique Dinand
    • 1
  • Anupam Sachdeva
    • 1
  • Sanghamitra Datta
    • 2
  • Sunita Bhalla
    • 3
  • Manas Kalra
    • 1
  • Chand Wattal
    • 2
  • Nita Radhakrishnan
    • 1
  1. 1.Pediatric Hematology, Oncology and BMT Unit, Department of Pediatrics, Institute of Child HealthSir Ganga Ram HospitalDelhiIndia
  2. 2.Department of Clinical Microbiology & ImmunologySir Ganga Ram HospitalNew DelhiIndia
  3. 3.Department of PathologySir Ganga Ram HospitalNew DelhiIndia

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