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Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples using automated sample preparation and real time PCR
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Reinhard B. Raggam
Published/Copyright:
December 15, 2009
Abstract
Background: Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples has gained significance in the routine diagnostic laboratory.
Methods: In this study, the analytical and clinical performance of the artus® EBV RG PCR kit in conjunction with automated sample preparation on the QIAsymphony® SP instrument was evaluated.
Results: When the accuracy of the new test system was tested, all results were found to be within ±0.5 log10 unit of the expected panel results. Determination of linearity showed a quasilinear curve over 4 log units. The lower limit of detection was determined to be 391 EBV DNA copies/mL in EDTA whole blood. The between day imprecision ranged from 18% to 66%, and the within run imprecision ranged from 11% to 50%. When clinical samples were tested and the results compared with those obtained with the routinely used easyMAG™ sample preparation and EBV R-gene™ test system, 60 samples tested positive and 31 samples tested negative by both assays. Nineteen samples were found to be positive using the QIAsymphony® sample preparation and artus® EBV RG PCR test system only, and no samples tested positive with the routinely used test system only.
Conclusions: The QIAsymphony® sample preparation and artus® EBV RG PCR test system is suitable for the detection and quantitation of EBV DNA in EDTA whole blood in the routine diagnostic laboratory.
Clin Chem Lab Med 2010;48:413–8.
Keywords: Epstein-Barr virus (EBV); molecular diagnostics; quantitation; real-time PCR; sample preparation
Received: 2009-9-26
Accepted: 2009-11-1
Published Online: 2009-12-15
Published in Print: 2010-03-01
©2010 by Walter de Gruyter Berlin New York
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Keywords for this article
Epstein-Barr virus (EBV);
molecular diagnostics;
quantitation;
real-time PCR;
sample preparation
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