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J Clin Virol. 2011 Mar;50(3):224-9. doi: 10.1016/j.jcv.2010.12.002. Epub 2010 Dec 24.

Multicenter comparative study of Epstein-Barr virus DNA quantification for virological monitoring in transplanted patients.

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  • 1Laboratory of Virology, INMI L. Spallanzani, Rome, Italy.

Abstract

BACKGROUND:

EBV-related post-transplant lymphoproliferative diseases are usually accompanied by increased EBV DNA in peripheral blood. Monitoring EBV DNAemia is the basis for weighing decisions regarding initiation of pre-emptive or anti-EBV-related tumor therapy. However, the definition of clinically relevant cut-off values is hampered by the lack of standardization in EBV DNA testing.

OBJECTIVES:

To estimate inter-laboratory variability and to evaluate the impact of different matrices in EBV DNA load determination in Italian laboratories involved in monitoring of virus infections in transplanted patients.

STUDY DESIGN:

Two different proficiency panels were distributed among seven centers: the first contained cell-associated and cell-free EBVs; the second was prepared by spiking both cell-associated and cell-free EBVs in EBV DNA-negative whole blood from EBV seropositive healthy donors. Samples were extracted and amplified with different methods. Intra-laboratory and inter-laboratory variabilities was evaluated.

RESULTS:

337 EBV DNA determinations were performed. Sensitivity was 100% for both panels, specificity was 100% for the first and 74% for the second panel, where whole blood was utilized as the matrix. Discrepant results in the second panel were restricted to samples containing low copy numbers. Quantification fell within ±0.5 log in 73% of the determinations. Values for cell-associated samples tended to be more heterogeneous than those obtained from cell-free samples. Good overall linearity was observed for each sample type; inter-laboratory variability ranged from 4.71% to 12.86%.

CONCLUSIONS:

The results of this multicenter study indicate that EBV DNAemia may be reliably quantified by different laboratories using a variety of commercial and in-house molecular assays.

Copyright © 2010 Elsevier B.V. All rights reserved.

PMID:
21185774
[PubMed - indexed for MEDLINE]
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