Abstract
BackgroundEpstein-Barr virus (EBV) DNA load monitoring is known to be useful for the diagnosis and monitoring of EBV-associated diseases. The aim of this study is to compare the performance of two real-time PCR assays for EBV DNA: a commercial kit as the Q-EBV Real-Time System (Q-EBV PCR, Amplimedical, Turin, Italy) and an in-house assay (EBV RQ-PCR).ResultsThe range of linearity and the degree of precision of the two assays were similar. The clinical sensitivity of Q-EBV PCR was higher for reference samples containing less than 1,000 EBV DNA copies/ml. The absolute quantitative results of the two methods were statistically correlated (R2 = 0.7789; p < 0.0001), with the systematic overestimation by EBV RQ-PCR possibly linked to different amplification efficiency in calibration standards.ConclusionBoth the commercial and the in-house assay may be appropriate for clinical use, but common standards are advisable for comparable absolute values, as these would improve the clinical utility of EBV DNA load measurement.
Highlights
Epstein-Barr virus (EBV) DNA load monitoring is known to be useful for the diagnosis and monitoring of EBV-associated diseases
Intraassay variability of the Q-EBV PCR and EBV RQ-PCR methods was determined by amplifying all samples in quadruplicate, whereas inter-assay variability was determined by amplifying the three dilutions in triplicate in four independent experiments
The quantification of EBV DNA load is useful for detecting viral reactivation, which increases the risk of post-transplant lymphoproliferative disease (PTLD) in immunosuppressed patients [16,17], and monitoring antiviral therapy
Summary
Epstein-Barr virus (EBV) DNA load monitoring is known to be useful for the diagnosis and monitoring of EBV-associated diseases. It has been estimated that the number of EBV-infected B cells is controlled in healthy individuals by EBV-specific immunity [1]. Active EBV infection is a strong risk factor for the development of post-transplant lymphoproliferative disease (PTLD) and AIDS-related lymphoma [6]. Quantitative molecular assays for the assessment of viral load have helped to describe and monitor EBV-related diseases. A range of different assay formats and protocols involving TaqMan probes [7,8] and fluorescence resonance energy transfer probes [9,10,11,12] have been reported and various in-house and commercial assays for (page number not for citation purposes)
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