scholarly journals Utility of Semiquantitative Polymerase Chain Reaction for Epstein‐Barr Virus to Measure Virus Load in Pediatric Organ Transplant Recipients with and without Posttransplant Lymphoproliferative Disease

2001 ◽  
Vol 33 (2) ◽  
pp. 145-150 ◽  
Author(s):  
Upton Allen ◽  
Diane Hebert ◽  
Martin Petric ◽  
Raymond Tellier ◽  
Dat Tran ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Epstein Barr Virus ◽  
Organ Transplant ◽  
Lymphoproliferative Disease ◽  
Transplant Recipients ◽  
Chain Reaction ◽  
Barr Virus ◽  
Posttransplant Lymphoproliferative Disease ◽  
Polymerase Chain ◽  
Epstein Barr

scholarly journals 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 ◽  
Vol 84 (3) ◽  
pp. 972-984 ◽  
Author(s):  
SA Riddler ◽  
MC Breinig ◽  
JL McKnight
Keyword(s):  
Epstein Barr Virus ◽  
Organ Transplant ◽  
Lymphoproliferative Disease ◽  
Nuclear Antigen ◽  
Solid Organ ◽  
Transplant Recipients ◽  
Barr Virus ◽  
Posttransplant Lymphoproliferative Disease ◽  
Solid Organ Transplant Recipients ◽  
Epstein Barr

Epstein-Barr virus (EBV)-associated posttransplant lymphoproliferative disease (PTLD) is an uncommon but potentially fatal complication of immunosuppression in solid-organ transplant recipients. A semiquantitative DNA polymerase chain reaction assay was developed to amplify a unique 269-bp region of the EBNA-1 gene in peripheral blood lymphocytes (PBL) using the primers described by Telenti et al (J Clin Microbiol 28:2187, 1990). Serial samples were studied from 23 transplant recipients, 12 of whom were diagnosed with PTLD. The majority of transplant recipients who were EBV seropositive at the time of transplant surgery and who did not develop PTLD (5 of 7, 71%) exhibited less than a 10-fold increase in the levels of EBV-infected PBL over the 0.1 to 5 EBV genomes/10(6) PBL observed in immunocompetent EBV seropositive controls. Transplant recipients who were seronegative at the time of transplantation and who underwent a primary EBV infection but did not develop PTLD exhibited a reduced capacity to control viremia because the levels of EBV-infected PBL were up to 400 times greater than the 1.0 to 50 EBV genomes/10(6) PBL observed in individuals undergoing acute infectious mononucleosis (Rocci et al: N Engl J Med 296:132, 1977). However, all transplant recipients who developed PTLD exhibited a marked elevation of EBV-infected PBL independent of their serologic state at the time of transplantation. Six of the 10 transplant recipients with PTLD exhibited > or = 300,000 EBV genomes/10(5) PBL, two exhibited 10,000 to 50,000 EBV-infected genomes/10(5) PBL, and one each exhibited 2,500 and 500 EBV genomes/10(5) PBL. However, the latter two samples were obtained 4 to 5 weeks after the diagnosis of PTLD and may reflect a decrease in viral load resulting from immunomodulation. Marked decreases in the levels of EBV nuclear antigen-1 (EBNA-1), EBNA-2, and EBNA-LP antibodies correlated with the increase in EBV-infected PBL. Hence, a quantitative difference in circulating EBV viral load and EBNA antibody levels is evident between transplant recipients with and without PTLD and may be useful as a noninvasive prognostic marker with which to monitor and/or predict the development of PTLD.

scholarly journals Semiquantitative Epstein-Barr Virus (EBV) Polymerase Chain Reaction for the Determination of Patients at Risk for EBV-Induced Lymphoproliferative Disease After Stem Cell Transplantation

Blood ◽  
1998 ◽  
Vol 91 (10) ◽  
pp. 3654-3661 ◽  
Author(s):  
Kenneth G. Lucas ◽  
Robert L. Burton ◽  
Sarah E. Zimmerman ◽  
Jinghong Wang ◽  
Kenneth G. Cornetta ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Stem Cell ◽  
T Cell ◽  
Epstein Barr Virus ◽  
Lymphoproliferative Disease ◽  
Chain Reaction ◽  
Barr Virus ◽  
Ebv Dna ◽  
Polymerase Chain ◽  
Epstein Barr

Abstract 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.

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