Automated quantification of Epstein-Barr Virus in whole blood of hematopoietic stem cell transplant patients using the Abbott m2000 system.
BACKGROUND
Accurate quantification of Epstein-Barr virus (EBV) load in blood is essential for the management of post-transplant lymphoproliferative disorders. The automation of DNA extraction and amplification may improve accuracy and reproducibility. We evaluated the EBV PCR Kit V1 with fully automated DNA extraction and amplification on the m2000 system (Abbott assay).


METHODS
Conversion factor between copies and international units (IU), lower limit of quantification, imprecision and linearity were determined in a whole blood (WB) matrix. Results from 339 clinical WB specimens were compared with a home-brew real-time PCR assay used in our laboratory (in-house assay).


RESULTS
The conversion factor between copies and IU was 3.22 copies/IU. The lower limit of quantification (LLQ) was 1000 copies/mL. Intra- and inter-assay coefficients of variation were 3.1% and 7.9% respectively for samples with EBV load higher than the LLQ. The comparison between Abbott assay and in-house assay showed a good concordance (kappa = 0.77). Loads were higher with the Abbott assay (mean difference = 0.62 log10 copies/mL).


CONCLUSIONS
The EBV PCR Kit V1 assay on the m2000 system provides a reliable and easy-to-use method for quantification of EBV DNA in WB.
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France. Electronic address: maud.salmona@sls.aphp.fr.
Maud
M
Salmona
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
Slim
S
Fourati
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
Linda
L
Feghoul
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
Catherine
C
Scieux
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
Aline
A
Thiriez
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
François
F
Simon
Univ Paris Diderot, Pres Sorbone Paris Cité, Biostatistics Department, APHP, Hôpital Saint-Louis, Paris, France.
Matthieu
M
Resche-Rigon
Univ Paris Diderot, Pres Sorbone Paris Cité, Inserm U941, APHP, Microbiology laboratory, Hôpital Saint-Louis, Paris, France.
Jérôme
J
LeGoff
D057205:Automation, Laboratory / Q000379:methods*; D001769:Blood / Q000821:virology*; D020031:Epstein-Barr Virus Infections / Q000821:virology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006801:Humans; D019562:Viral Load / Q000379:methods*
Automated extraction and quantification; Epstein-Barr Virus; Hematopoietic stem cell transplantation; Real-time PCR; Whole-blood