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Comparative Study
. 2008 Jan;46(1):157-63.
doi: 10.1128/JCM.01252-07. Epub 2007 Nov 7.

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

Affiliations
Comparative Study

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

R T Hayden et al. J Clin Microbiol. 2008 Jan.

"VSports手机版" 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 VSports手机版. 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/microl) 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. .

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Figures

FIG. 1.
FIG. 1.
Comparison of calibration curves for all test panel runs.
FIG. 2.
FIG. 2.
Comparison of calibration curves by laboratory, comparing all test panel runs.
FIG. 3.
FIG. 3.
Variation in viral load values for patient samples. Each point gives the result from a specific laboratory for the indicated panel and sample number. The horizontal line at y = 1 corresponds to the threshold used to qualitatively interpret the findings as positive or negative. The numbers at the bottom of each plot indicate the number of laboratories that did not report a CT value and interpreted the result as negative (zeroes are not shown).
FIG. 4.
FIG. 4.
Consistency of results for negative and low viral load samples. Each solid line shows the results of a negative or low viral sample for one panel across labs. The plot includes all instances with a mean viral load less than or equal to 2 on the log-scale. The results deemed negative by the lab are shown as −4 on this plot. A dashed horizontal line at y = 1 is included for purposes of qualitative interpretation.

References

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