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METHODOLOGY Open Access Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing

By Two Ebv Dna Targets, Sybr Green I Dye, Meav-lang J Lay, Robyn M Lucas, Mala Ratnamohan, Janette Taylor, Anne-louise Ponsonby, Dominic E Dwyer and The Ausimmune Investigator Group (aig

Abstract

Background: Reactivation of Epstein-Barr virus (EBV) infection may cause serious, life-threatening complications in immunocompromised individuals. EBV DNA is often detected in EBV-associated disease states, with viral load believed to be a reflection of virus activity. Two separate real-time quantitative polymerase chain reaction (QPCR) assays using SYBR Green I dye and a single quantification standard containing two EBV genes, Epstein-Barr nuclear antigen-1 (EBNA-1) and BamHI fragment H rightward open reading frame-1 (BHRF-1), were developed to detect and measure absolute EBV DNA load in patients with various EBV-associated diseases. EBV DNA loads and viral capsid antigen (VCA) IgG antibody titres were also quantified on a population sample. Results: EBV DNA was measurable in ethylenediaminetetraacetic acid (EDTA) whole blood, peripheral blood mononuclear cells (PBMCs), plasma and cerebrospinal fluid (CSF) samples. EBV DNA loads were detectable from 8.0 × 10 2 to 1.3 × 10 8 copies/ml in post-transplant lymphoproliferative disease (n = 5), 1.5 × 10 3 to 2.0 × 10 5 copies/ml in infectious mononucleosis (n = 7), 7.5 × 10 4 to 1.1 × 10 5 copies/ml in EBV-associated haemophagocytic syndrome (n = 1), 2.0 × 10 2 to 5.6 × 10 3 copies/ml in HIV-infected patients (n = 12), and 2.0 × 10 2 to 9.1 × 10 4 copies/ml in the population sample (n = 218). EBNA-1 and BHRF-1 DNA were detected in 11.0 % and 21.6 % of the population sample respectively. There was a modest correlation between VCA IgG antibody titre and BHRF-1 DNA load (rho = 0.13, p = 0.05) but no

Year: 2013
OAI identifier: oai:CiteSeerX.psu:10.1.1.352.1110
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