• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Jun 1997; 35(6): 1612–1615.
PMCID: PMC229802

Use of quantitative competitive PCR to measure Epstein-Barr virus genome load in the peripheral blood of pediatric transplant patients with lymphoproliferative disorders.


A quantitative competitive PCR (QC-PCR) assay for Epstein-Barr virus (EBV) has been developed to provide accurate measurement of EBV genome load in pediatric transplant recipients at risk for developing posttransplant lymphoproliferative disorder (PTLD). The assay quantifies between 8 and 5,000 copies of the EBV genome in 10(5) lymphocytes after a 30-cycle amplification reaction. For 14 pediatric patients diagnosed with PTLD, the median EBV genome load was 4,000, and 13 of the 14 patients had values of >500 copies per 10(5) lymphocytes. Only 3 of 12 control transplant recipients not diagnosed with PTLD had detectable viral genome loads (median value, 40). This median was calculated by using the highest value obtained by PCR testing on each of these patients posttransplantation. PCR values of >500 copies per 10(5) lymphocytes appear to correlate with a diagnosis of PTLD. By a modified protocol, the EBV genome copy number in latently infected adults was estimated to be <0.1 copy per 10(5) lymphocytes.

Full Text

The Full Text of this article is available as a PDF (358K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Cen H, Breinig MC, Atchison RW, Ho M, McKnight JL. Epstein-Barr virus transmission via the donor organs in solid organ transplantation: polymerase chain reaction and restriction fragment length polymorphism analysis of IR2, IR3, and IR4. J Virol. 1991 Feb;65(2):976–980. [PMC free article] [PubMed]
  • Chen F, Zou JZ, di Renzo L, Winberg G, Hu LF, Klein E, Klein G, Ernberg I. A subpopulation of normal B cells latently infected with Epstein-Barr virus resembles Burkitt lymphoma cells in expressing EBNA-1 but not EBNA-2 or LMP1. J Virol. 1995 Jun;69(6):3752–3758. [PMC free article] [PubMed]
  • Craig FE, Gulley ML, Banks PM. Posttransplantation lymphoproliferative disorders. Am J Clin Pathol. 1993 Mar;99(3):265–276. [PubMed]
  • de-Thé G, Geser A, Day NE, Tukei PM, Williams EH, Beri DP, Smith PG, Dean AG, Bronkamm GW, Feorino P, et al. Epidemiological evidence for causal relationship between Epstein-Barr virus and Burkitt's lymphoma from Ugandan prospective study. Nature. 1978 Aug 24;274(5673):756–761. [PubMed]
  • Gutierrez MI, Bhatia K, Magrath I. Replicative viral DNA in Epstein-Barr virus associated Burkitt's lymphoma biopsies. Leuk Res. 1993 Mar;17(3):285–289. [PubMed]
  • Hanto DW, Gajl-Peczalska KJ, Frizzera G, Arthur DC, Balfour HH, Jr, McClain K, Simmons RL, Najarian JS. Epstein-Barr virus (EBV) induced polyclonal and monoclonal B-cell lymphoproliferative diseases occurring after renal transplantation. Clinical, pathologic, and virologic findings and implications for therapy. Ann Surg. 1983 Sep;198(3):356–369. [PMC free article] [PubMed]
  • Kenagy DN, Schlesinger Y, Weck K, Ritter JH, Gaudreault-Keener MM, Storch GA. Epstein-Barr virus DNA in peripheral blood leukocytes of patients with posttransplant lymphoproliferative disease. Transplantation. 1995 Sep 27;60(6):547–554. [PubMed]
  • Khanna R, Burrows SR, Moss DJ. Immune regulation in Epstein-Barr virus-associated diseases. Microbiol Rev. 1995 Sep;59(3):387–405. [PMC free article] [PubMed]
  • Klein G. Specific chromosomal translocations and the genesis of B-cell-derived tumors in mice and men. Cell. 1983 Feb;32(2):311–315. [PubMed]
  • Klein G. Epstein-Barr virus strategy in normal and neoplastic B cells. Cell. 1994 Jun 17;77(6):791–793. [PubMed]
  • Kovalic D, Kwak JH, Weisblum B. General method for direct cloning of DNA fragments generated by the polymerase chain reaction. Nucleic Acids Res. 1991 Aug 25;19(16):4560–4560. [PMC free article] [PubMed]
  • Lager DJ, Burgart LJ, Slagel DD. Epstein-Barr virus detection in sequential biopsies from patients with a posttransplant lymphoproliferative disorder. Mod Pathol. 1993 Jan;6(1):42–47. [PubMed]
  • Masucci MG. Viral immunopathology of human tumors. Curr Opin Immunol. 1993 Oct;5(5):693–700. [PubMed]
  • Nalesnik MA, Jaffe R, Starzl TE, Demetris AJ, Porter K, Burnham JA, Makowka L, Ho M, Locker J. The pathology of posttransplant lymphoproliferative disorders occurring in the setting of cyclosporine A-prednisone immunosuppression. Am J Pathol. 1988 Oct;133(1):173–192. [PMC free article] [PubMed]
  • Penn I, Hammond W, Brettschneider L, Starzl TE. Malignant lymphomas in transplantation patients. Transplant Proc. 1969 Mar;1(1):106–112. [PMC free article] [PubMed]
  • Preiksaitis JK, Diaz-Mitoma F, Mirzayans F, Roberts S, Tyrrell DL. Quantitative oropharyngeal Epstein-Barr virus shedding in renal and cardiac transplant recipients: relationship to immunosuppressive therapy, serologic responses, and the risk of posttransplant lymphoproliferative disorder. J Infect Dis. 1992 Nov;166(5):986–994. [PubMed]
  • Randhawa PS, Jaffe R, Demetris AJ, Nalesnik M, Starzl TE, Chen YY, Weiss LM. The systemic distribution of Epstein-Barr virus genomes in fatal post-transplantation lymphoproliferative disorders. An in situ hybridization study. Am J Pathol. 1991 Apr;138(4):1027–1033. [PMC free article] [PubMed]
  • Rea D, Fourcade C, Leblond V, Rowe M, Joab I, Edelman L, Bitker MO, Gandjbakhch I, Suberbielle C, Farcet JP, et al. Patterns of Epstein-Barr virus latent and replicative gene expression in Epstein-Barr virus B cell lymphoproliferative disorders after organ transplantation. Transplantation. 1994 Aug 15;58(3):317–324. [PubMed]
  • Riddler SA, Breinig MC, McKnight JL. Increased levels of circulating Epstein-Barr virus (EBV)-infected lymphocytes and decreased EBV nuclear antigen antibody responses are associated with the development of posttransplant lymphoproliferative disease in solid-organ transplant recipients. Blood. 1994 Aug 1;84(3):972–984. [PubMed]
  • Rooney CM, Loftin SK, Holladay MS, Brenner MK, Krance RA, Heslop HE. Early identification of Epstein-Barr virus-associated post-transplantation lymphoproliferative disease. Br J Haematol. 1995 Jan;89(1):98–103. [PubMed]
  • Savoie A, Perpête C, Carpentier L, Joncas J, Alfieri C. Direct correlation between the load of Epstein-Barr virus-infected lymphocytes in the peripheral blood of pediatric transplant patients and risk of lymphoproliferative disease. Blood. 1994 May 1;83(9):2715–2722. [PubMed]
  • Starzl TE, Nalesnik MA, Porter KA, Ho M, Iwatsuki S, Griffith BP, Rosenthal JT, Hakala TR, Shaw BW, Jr, Hardesty RL, et al. Reversibility of lymphomas and lymphoproliferative lesions developing under cyclosporin-steroid therapy. Lancet. 1984 Mar 17;1(8377):583–587. [PMC free article] [PubMed]
  • Swerdlow SH. Post-transplant lymphoproliferative disorders: a morphologic, phenotypic and genotypic spectrum of disease. Histopathology. 1992 May;20(5):373–385. [PubMed]
  • Thorley-Lawson DA, Mann KP. Early events in Epstein-Barr virus infection provide a model for B cell activation. J Exp Med. 1985 Jul 1;162(1):45–59. [PMC free article] [PubMed]
  • Thorley-Lawson DA. Immunological responses to Epstein-Barr virus infection and the pathogenesis of EBV-induced diseases. Biochim Biophys Acta. 1989 Feb;948(3):263–286. [PubMed]
  • Tierney RJ, Steven N, Young LS, Rickinson AB. Epstein-Barr virus latency in blood mononuclear cells: analysis of viral gene transcription during primary infection and in the carrier state. J Virol. 1994 Nov;68(11):7374–7385. [PMC free article] [PubMed]
  • Wagner HJ, Bein G, Bitsch A, Kirchner H. Detection and quantification of latently infected B lymphocytes in Epstein-Barr virus-seropositive, healthy individuals by polymerase chain reaction. J Clin Microbiol. 1992 Nov;30(11):2826–2829. [PMC free article] [PubMed]
  • Weiss LM, Movahed LA. In situ demonstration of Epstein-Barr viral genomes in viral-associated B cell lymphoproliferations. Am J Pathol. 1989 Mar;134(3):651–659. [PMC free article] [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...