Performance Characteristics of Two Real-Time PCR Assays for the Quantification of Epstein-Barr Virus DNA

Abstract

We compared the performance of a laboratory-developed 5'-nuclease real-time polymerase chain reaction assay and a commercial assay (LightCycler, Roche Diagnostics, Indianapolis, IN) for quantification of Epstein-Barr virus (EBV) DNA. Using standards provided by the manufacturer, the LightCycler assay was linear from 100 to 1 million copies per reaction. Based on dilution of a plasmid containing the amplicon, the laboratory-developed assay was linear from 22 to 45 million copies per reaction. Both assays detected 0.5 copies of genomic EBV DNA per reaction; both showed good reproducibility with coefficients of variation from 0.3% to 2.4% for the LightCycler and 1.8% to 5.1% for the laboratory-developed assay. For 31 whole blood specimens with measurable values in both assays, the viral load values obtained with the LightCycler averaged 2.3-fold higher than those obtained with the laboratory-developed assay. Testing 30 matched whole blood and plasma samples in the laboratory-developed assay showed whole blood viral load values averaged 10-fold higher than those for plasma. The LightCycler and laboratory-developed assays are sensitive and reproducible with broad linear ranges. Further clinical evaluation is needed to determine the viral load cutoff that is predictive of posttransplantation lymphoproliferative disorders.

A Global View of the Oncogenic Landscape in Nasopharyngeal Carcinoma: An Integrated Analysis at the Genetic and Expression Levels

“…The samples and their origins are listed in Table 1. Samples were tested for EBV DNA-positivity by PCR on extracted DNA [59] and for the expression of EBV encoded genes by RT-PCR as described [60] using the primers listed in Table S8.…”

JC virus DNA in the peripheral blood of renal transplant patients: A 1‐year prospective follow‐up in France

“…HCMV DNA was quantified on a Light Cycler 1.0 TM (Roche Diagnostics), as described previously [Mengelle et al, 2003]. EBV DNA was quantified using a Light Cycler EBV Quantification Kit1 (Roche Diagnostics), according to the manufacturer's instructions [Gulley et al, 2006;Hill et al, 2006;Kozic et al, 2006]. BKV DNA was quantified on a Light Cycler 2.0 TM (Roche Diagnostics), as described previously [Basse et al, 2007].…”

Levels of oxidative stress biomarkers and bone resorption regulators in apical periodontitis lesions infected by Epstein–Barr virus

“…Epstein-Barr virus in apical periodontitis Jakovljevic et al fluorescence labelled at the 5 0 end with 6-carboxyfluorescein (FAM) as the reporter dye and at the 3 0 end with 6-carboxy-tetramethyl-rhodamide (TAMRA) dye (5 0 -FAM TCC TGC AGC TAT TTC TGG TCG CAT CA TAMRA-3 0 ). Details on the sensitivity and specificity of the sets of primers have been described previously (Hill et al 2006).…”