Evaluation of a commercial real-time PCR assay for quantitation of Epstein-Barr virus DNA in different groups of patients

• Published in: Journal of virological methods 2006-08, Vol.135 (2), p.263-268
• Main Authors: Kozić, Sanja, Vince, Adriana, Beš, Janja Iščić, Rode, Oktavija Đaković, Lepej, Snježana Židovec, Poljak, Mario, Bozic, Michael, Kessler, Harald H.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; DNA, Viral - analysis; Epstein-Barr virus; Fundamental and applied biological sciences. Psychology; Herpesvirus 4, Human - genetics; Human immunodeficiency virus; Humans; Immunosupression; Microbiology; Polymerase Chain Reaction - methods; Quantitation; Reagent Kits, Diagnostic; Real-time PCR; Techniques used in virology; Viral load; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2006.03.013

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.