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Validation of an Epstein-Barr viral load assay using the QIAGEN Artus EBV TM PCR analyte-specific reagent
Arshad N Ahsanuddin1, M Craig Standish, Angela M Caliendo
1Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Emory University Hospital, Atlanta, GA 30322, USA.
We describe the validation of a test for the quantification of Epstein-Barr virus (EBV) DNA (viral load) using the Artus EBV TM PCR analyte-specific reagent (ASR; QIAGEN Hamburg, Hamburg, Germany). A dilution series demonstrated a limit of detection of 2.25 log(10) copies/mL (>95% positivity rate). The limit of quantification was 3.90 log(10) copies/mL based on an SD of less than 0.15. The assay was linear from 2.17 to 6.2 log(10) copies/mL. Low (3.70 log(10) copies/mL) and high (5.40 log(10) copies/mL) patient samples had coefficients of variation (CVs) of 2.0% and 1.4%, respectively. The cycle thresholds of 4 points used to generate the standard curve had CVs ranging from 0.8% to 1.6%. A comparison of 35 matched samples showed a small positive bias (0.35 log(10) copies/mL) for the Artus ASR relative to a laboratory-developed EBV viral load assay targeting the Bam H1-W region of the EBV genome.
We describe the validation of a test for the quantification of Epstein-Barr virus (EBV) DNA (viral load) using the Artus EBV TM PCR analyte-specific reagent (ASR; QIAGEN Hamburg, Hamburg, Germany). A dilution series demonstrated a limit of detection of 2.25 log(10) copies/mL (>95% positivity rate). The limit of quantification was 3.90 log(10) copies/mL based on an SD of less than 0.15. The assay was linear from 2.17 to 6.2 log(10) copies/mL. Low (3.70 log(10) copies/mL) and high (5.40 log(10) copies/mL) patient samples had coefficients of variation (CVs) of 2.0% and 1.4%, respectively. The cycle thresholds of 4 points used to generate the standard curve had CVs ranging from 0.8% to 1.6%. A comparison of 35 matched samples showed a small positive bias (0.35 log(10) copies/mL) for the Artus ASR relative to a laboratory-developed EBV viral load assay targeting the Bam H1-W region of the EBV genome.
