Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples using automated sample preparation and real time PCR.
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 log(10) 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.
Raggam
Reinhard B
RB
Research Unit Molecular Diagnostics, Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Wagner
Jasmin
J
Bozic
Michael
M
Michelin
Birgit D A
BD
Hammerschmidt
Sandra
S
Homberg
Cordula
C
Kessler
Harald H
HH
D001331:Automation; D004279:DNA, Viral / Q000097:blood*; D004492:Edetic Acid / Q000737:chemistry*; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D004492:Edetic Acid