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Semiquantitative Epstein-Barr Virus (EBV) Polymerase Chain
Reaction for the Determination of Patients at Risk for
EBV-Induced Lymphoproliferative Disease After Stem Cell
Transplantation
Kenneth G. Lucas,
Robert L. Burton,
Sarah E. Zimmerman,
Jinghong Wang,
Kenneth G. Cornetta,
Kent A. Robertson,
Chao H. Lee, and
David J. Emanuel
From the Department of Pediatrics, Stem Cell Transplantation Program,
Riley Hospital for Children; The Department of Medicine, Bone Marrow
Transplantation Program; and the Department of Pathology and Laboratory
Medicine, Indiana University Medical Center, Indianapolis, IN.
Epstein-Barr virus-induced lymphoproliferative disease (EBV-LPD) is
a serious and potentially fatal complication after allogeneic stem cell
transplantation (SCT). To evaluate levels of EBV DNA in SCT patients, a
semiquantitative polymerase chain reaction (PCR) assay was developed.
DNA was extracted from peripheral blood leukocytes and diluted, and PCR
was performed by using a primer set specific for a well-conserved
sequence of the internal repeat 1 region of the EBV genome. Forty-one
SCT patients were screened with this method. Thirty-seven patients
received allogeneic transplants, of which 18 were T-cell-depleted
marrow. Four additional patients received autologous SCT, one of which
was T-cell depleted. The mean time of follow-up by EBV PCR was 147 days
(range, 47 to 328 days) posttransplant. The range of EBV copies/µg
DNA from normal EBV sero-positive donors was 40 to 4,000. Seven
patients had  40,000 copies of EBV DNA/µg DNA, all of whom were
recipients of T-cell-depleted SCT. Five of the seven patients with
elevated levels of EBV DNA developed EBV-LPD. Four of these five
patients with EBV-LPD had elevated levels of EBV DNA from 1 to 8 weeks
before diagnosis. Two patients with EBV-LPD had normal levels of EBV
DNA, and two patients with 40,000 copies EBV/µg DNA did not
develop EBV-LPD. In one patient, clinical resolution of disease
correlated with a decrease in EBV DNA and an increase in the level of
EBV-specific cytotoxic T-cell precursors. These data indicate that the
measurement of EBV viral load with semiquantitative PCR is useful in
detecting EBV-LPD in high-risk patients before the onset of clinical
symptoms. Because not all patients with elevated levels of EBV DNA
develop EBV-LPD, semiquantitative PCR results cannot substitute for
clinical, radiographic, and pathological confirmation of this
diagnosis.
Blood, Vol. 91 No. 10 (May 15), 1998:
pp. 3654-3661
© 1998 by The American Society of Hematology.
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