Multicenter comparison of different real-time PCR assays for quantitative detection of Epstein-Barr virus

R. T. Hayden, K. M. Hokanson, S. B. Pounds, M. J. Bankowski, S. W. Belzer, J. Carr, D. Diorio, M. S. Forman, Y. Joshi, D. Hillyard, R. L. Hodinka, M. N. Nikiforova, Charlotte A Romain, J. Stevenson, A. Valsamakis, Henry H Balfour

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93 Scopus citations

Abstract

Quantification of Epstein-Barr virus (EBV) in peripheral blood is important for the diagnosis and management of serious EBV diseases, including posttransplant lymphoproliferative disorder. A variety of PCR-based methods are currently in use; however, there is little information on their comparability. This study assessed the relative performance of different quantitative assays. A multicenter comparative study was performed at eight sites using three panels consisting of serial dilutions of quantified EBV DNA and extracts from a total of 19 whole-blood specimens. Samples were distributed and tested blindly. Instrumentation, probe chemistries, amplification targets, and other test-related aspects varied considerably between laboratories. Each laboratory's calibration curve indicated strong evidence of a consistent log-linear relationship between viral load and cycle threshold, suggesting that intralaboratory tracking of a given patient would yield similar relative quantitative trends among the participating test sites. There was strong concordance among laboratories with respect to qualitative test results; however, marked quantitative discordance was seen. For most samples, the across-laboratory interquartile range of the reported viral load (in copies/μl) was roughly 0.6 log-units, and for one sample the overall range was approximately 4.2 log-units. While intralaboratory tracking of patients may yield similar results, these data indicate a need for caution when attempting to compare clinical results obtained at different institutions and suggest the potential value to be gained by more standardized testing methodology.

Original languageEnglish (US)
Pages (from-to)157-163
Number of pages7
JournalJournal of clinical microbiology
Volume46
Issue number1
DOIs
StatePublished - Jan 1 2008

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    Hayden, R. T., Hokanson, K. M., Pounds, S. B., Bankowski, M. J., Belzer, S. W., Carr, J., Diorio, D., Forman, M. S., Joshi, Y., Hillyard, D., Hodinka, R. L., Nikiforova, M. N., Romain, C. A., Stevenson, J., Valsamakis, A., & Balfour, H. H. (2008). Multicenter comparison of different real-time PCR assays for quantitative detection of Epstein-Barr virus. Journal of clinical microbiology, 46(1), 157-163. https://doi.org/10.1128/JCM.01252-07