Measurement of Epstein-Barr Virus DNA Loads in Whole Blood and Plasma by TaqMan PCR and in Peripheral Blood Lymphocytes by Competitive PCR

Wadowsky, Robert M.; Laus, Stella; Green, Michael; Webber, Steven A.; Rowe, David

doi:10.1128/jcm.41.11.5245-5249.2003

Cited by 101 publications

(76 citation statements)

References 11 publications

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“…Several studies reported that quantifying cell-free EBV DNA predicted the development of PTLD [59,68,69]. However, serum or plasma samples lack cell-associated virus and therefore plasma loads are not correlated with PBMC values [70]. Stevens et al reported that the increased EBV DNA loads in PTLD patients were restricted to the cellular compartment, as parallel serum samples were below the cut-off value [67].…”

Section: Post-transplant Lymphoproliferative Disordermentioning

confidence: 99%

“…More recently, unfractionated whole blood has been used because whole blood can be obtained readily and contains all blood compartments that may harbour EBV. There have been several reports that whole blood is better than plasma/serum when testing PTLD patients [67,[70][71][72][73]. Based on these observations, whole blood is the preferred specimen for PTLD Table 3), although more thorough studies are needed to resolve this controversy.…”

Section: Post-transplant Lymphoproliferative Disordermentioning

confidence: 99%

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Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Kimura

Ito

Suzuki

et al. 2008

Reviews in Medical Virology

139

142

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Because Epstein-Barr virus (EBV) is ubiquitous and persists latently in lymphocytes, simply detecting EBV is insufficient to diagnose EBV-associated diseases. Therefore, measuring the EBV load is necessary to diagnose EBVassociated diseases and to explore EBV pathogenesis. Due to the diverse biology of EBV, the significance of measuring EBV DNA and the optimal type of specimen differ among EBV-associated diseases. Recent advances in molecular technology have enabled the EBV genome to be quantitated rapidly and accurately. Real-time polymerase chain reaction (PCR) is a rapid and reliable method to quantify DNA and is widely used not only as a diagnostic tool, but also as a management tool for EBV-associated diseases. However, each laboratory currently measures EBV load with its own ''homebrew'' system, and there is no consensus on sample type, sample preparation protocol, or assay units. The EBV real-time PCR assay system must be standardised for large-scale studies and international comparisons.

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Section: Post-transplant Lymphoproliferative Disordermentioning

confidence: 99%

Section: Post-transplant Lymphoproliferative Disordermentioning

confidence: 99%

Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Kimura

Ito

Suzuki

et al. 2008

Reviews in Medical Virology

139

142

View full text Add to dashboard Cite

show abstract

“…Subsequent post-transplant EBV serology was positive in all 11 patients who had clinical data available (Table 1). Whole-blood EBV viral loads estimated by quantitative PCR testing 21 are reported by our institution into clinically relevant quartiles, with quartile groups III (16,000-799,999 copies/mL) and IV (>800,000 copies/mL) historically associated with increased risk of either having or developing an EBV-PTLD. Of the 8 patients who had quantitative EBV viral-load testing 1 month before their diagnosis, 6 of them had high levels, according to our institution's clinical quartiles III and IV (Table 1).…”

Section: Flow Cytometrymentioning

confidence: 99%

Post‐transplant Burkitt lymphoma is a more aggressive and distinct form of post‐transplant lymphoproliferative disorder

Picarsic

Jaffe

Mazariegos

et al. 2011

Cancer

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BACKGROUND:Although the literature reports a low incidence of Burkitt lymphoma (BL) as a post‐transplant lymphoproliferative disorder (PTLD), this entity appears to be different from other monomorphic PTLDs (M‐PTLDs), both in its aggressive clinical presentation and its distinct pathologic profile.METHODS:Patients with BL, diagnosed in the post‐transplant setting, (patients aged ≤18 years) were retrieved from the pathology archives at Children's Hospital of Pittsburgh of the University of Pittsburgh Medical Center from 1982 to 2010. Clinical outcomes were obtained along with pathologic review.RESULTS:Twelve patients with pediatric BL in the post‐transplant setting (9 boys, 3 girls) were retrieved. The patients displayed a monomorphic population of small to intermediate‐sized, noncleaved, lymphoid elements with a “starry‐sky” pattern. The immunophenotype for patients available to the study was CD20+ (n = 9/10), CD10+ (n = 8/8), bcl‐6+ (n = 11/11), with a near 100% Ki‐67/MIB‐1 proliferation index (n = 7/7), and negative for TdT (n = 7/7). Most pretransplant Epstein‐Barr virus titers were negative (n = 8/10), with post‐transplant titers positive in all tested patients (n = 11), and with positive Epstein‐Barr–encoded RNA in situ hybridization in most cases (n = 9/11). The median time from transplantation to diagnosis was 52 months (range, 6‐107 months). Nine patients were currently alive after immediate antineoplastic chemotherapy, with median disease‐free time of 93 months from diagnosis (range, 2‐199 months).CONCLUSIONS:BL‐PTLD had a higher Epstein‐Barr virus incidence compared with sporadic and immunodeficiency‐associated BL and represented a distinct monomorphic PTLD. Although some M‐PTLDs can be managed less aggressively with decreased immunosuppression alone, immediate lymphoma‐specific chemotherapy was associated with a favorable outcome and was strongly recommended. Cancer 2011;. © 2011 American Cancer Society.

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“…Previous studies have shown a good correlation when comparing EBV DNA load in whole blood and plasma [Stevens et al, 2001;Wadowsky et al, 2003] or in whole blood and separated leukocytes [Razonable et al, 2002;Hakim et al, 2007]. Mean viral loads reported in copies/10 6 PBLs are 0.4 log 10 units lower than those reported per ml of whole blood.…”

Section: Discussionmentioning

confidence: 92%

Cellular normalization of viral DNA loads on whole blood improves the clinical management of cytomegalovirus or Epstein Barr virus infections in the setting of pre‐emptive therapy

Bressollette-Bodin

Coste‐Burel

Besse

et al. 2008

Journal of Medical Virology

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Two quantitative duplex real-time PCR assays were developed for co-amplification of human albumin and cytomegalovirus (CMV) or Epstein Barr virus (EBV) genes after automated extraction on whole blood, and compared two units for expressing viral DNA loads (copies per ml of blood or per 10 6 peripheral blood leukocytes (PBLs)) on 1,138 positive samples. Both PCRs were characterized by high sensitivity, reproducibility, and linear range. Automated extraction by a MagNA Pure LC Instrument was shown to be more efficient when peripheral blood cell count was inferior to 5 Â 10 9 PBLs/L. Albumin coamplification allows the detection of PCR inhibitors and normalization of viral load according to the number of cells calculated in the sample. The two ways of expressing viral load results were highly correlated, but quantitative differences varied in relation to variations of blood cell count. As these two viruses are highly cell associated, viral loads can be underestimated in patients with leucopenia. In the setting of pre-emptive strategies during CMV infection, the units in which results are expressed can influence clinical management, as illustrated in this article.

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Measurement of Epstein-Barr Virus DNA Loads in Whole Blood and Plasma by TaqMan PCR and in Peripheral Blood Lymphocytes by Competitive PCR

Cited by 101 publications

References 11 publications

Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Measuring Epstein–Barr virus (EBV) load: the significance and application for each EBV‐associated disease

Post‐transplant Burkitt lymphoma is a more aggressive and distinct form of post‐transplant lymphoproliferative disorder

Cellular normalization of viral DNA loads on whole blood improves the clinical management of cytomegalovirus or Epstein Barr virus infections in the setting of pre‐emptive therapy

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