Semiquantitative Epstein-Barr virus (EBV) polymerase chain reaction for the determination of patients at risk for EBV-induced lymphoproliferative disease after stem cell transplantation.
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/microg DNA from normal EBV sero-positive donors was 40 to 4,000. Seven patients had >/=40,000 copies of EBV DNA/microg 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/microg 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.
Lucas
K G
KG
Department of Pediatrics, Stem Cell Transplantation Program, Riley Hospital for Children, Indianapolis, IN 46202-5225, USA.
Burton
R L
RL
Zimmerman
S E
SE
Wang
J
J
Cornetta
K G
KG
Robertson
K A
KA
Lee
C H
CH
Emanuel
D J
DJ
D000293:Adolescent; D000328:Adult; D000368:Aged; D000961:Antilymphocyte Serum / Q000009:adverse effects; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000097:blood*; D005260:Female; D005500:Follow-Up Studies; D006086:Graft vs Host Disease / Q000209:etiology; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000635:transmission / Q000821:virology; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007938:Leukemia / Q000150:complications / Q000628:therapy; D017708:Leukocyte Transfusion / Q000009:adverse effects; D007962:Leukocytes / Q000821:virology; D008212:Lymphocyte Depletion / Q000009:adverse effects; D008232:Lymphoproliferative Disorders / Q000453:epidemiology / Q000209:etiology / Q000821:virology*; D008297:Male; D009190:Myelodysplastic Syndromes / Q000150:complications / Q000628:therapy; D016133:Polymerase Chain Reaction / Q000379:methods*; D012306:Risk; D013601:T-Lymphocytes; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000635:transmission / Q000821:virology
D000961:Antilymphocyte Serum; D004279:DNA, Viral