Evaluation of a commercial real-time PCR assay for quantitation of Epstein-Barr virus DNA in different groups of patients.
The aim of this study was to evaluate the performance of a molecular assay for quantitation of Epstein-Barr virus (EBV) DNA based on real-time PCR, and to determine EBV DNA levels in EDTA whole blood samples derived from different groups of patients. Following a manual DNA extraction protocol, real-time PCR was performed using the LightCycler EBV Quantification Kit, which demonstrated sufficient accuracy and linearity. Coefficients of variations were found to be between 6 and 42% and 5 and 34%, respectively, for inter-assay and intra-assay variations. In clinical specimens, EBV DNA was detected in all patients with acute EBV infection (n=34), in one of 25 adults with past EBV infection, in 16 out of 25 (64%) anti-HIV antibody positive persons, in 10 out of 25 (40%) solid organ transplant recipients, and in none of the 23 infants without history of EBV infection. When EBV DNA levels in positive specimens were compared between different groups, statistically significant differences were not found. The LightCycler EBV Quantification Kit was found to be useful for determination of EBV DNA levels in EDTA whole blood.
Kozić
Sanja
S
University Hospital for Infectious Diseases, Dr. Fran Mihaljević, Zagreb, Croatia. skozic@bfm.hr
Vince
Adriana
A
Bes
Janja Iscić
JI
Rode
Oktavija Daković
OD
Lepej
Snjezana Zidovec
SZ
Poljak
Mario
M
Bozic
Michael
M
Kessler
Harald H
HH
D004279:DNA, Viral / Q000032:analysis*; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic