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Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples
Published on: June 30, 2018
Comparison of two highly automated DNA extraction systems for quantifying Epstein-Barr virus in whole blood
C Mengelle1, F Legrand-Abravanel, J-M Mansuy
1Department of Virology, CH Toulouse, France. mengelle.c@chu-toulouse.fr
Background:
Optimal automated molecular methods are needed to monitor Epstein-Barr virus (EBV) infections in transplant recipients.
Objectives:
To compare the extraction of EBV DNA from whole blood using the COBAS Ampliprep and the MagNA Pure instruments (Roche) for quantifying EBV DNA by real-time PCR.
Study Design:
EBV DNA content was determined on clinical samples extracted by both systems.
Results:
The detection limit was 2.16log(10)copies/mL using the COBAS Ampliprep extraction system. Specificity was 100% and we saw no cross-contamination. Extraction was linear from 2.60 to 5.60log(10)copies/mL. The intra-assay variation was 1.91% for 3.60, 2% for 4.60 and 4.51% for 5.60log(10)copies/mL; inter-assay variation was 4.88%. Sixty-six samples were tested: 26 were positive and 28 were negative by both methods. One sample was MagNA Pure positive/COBAS Ampliprep negative (virus load 3.15log(10)copies/mL) and 10 samples were MagNA Pure negative/COBAS Ampliprep positive (virus loads from 1.59 to 3.51log(10)copies/mL) (P<0.0001). Both methods gave similar quantitative results (average difference 0.07log(10)copies/mL) which were well correlated (r=0.73, P<0.001).
Conclusions:
The COBAS Ampliprep extraction system is comparable to the MagNA Pure and offers a high reliability for extracting EBV DNA from whole blood.
