Quantitative Epstein-Barr virus (EBV) DNA determinations are obtained by LightCycler PCR.
Viral DNA is extracted by the MagNA Pure automated instrument (Roche Applied Science) from whole blood. LightCycler PCR primers and probes have been designed to detect target EBV DNA. Amplified product is detected by a fluorescence assay (fluorescence resonance energy transfer: FRET). Quantitative results are obtained by incorporating known copy levels (5 standards) of EBV DNA (plasmid) into the assay together with the sample. LightCycler instrument software calculates the level of EBV DNA (copy/mL) in the sample from a standard curve of the quantitative levels of the reference plasmid EBV DNA.(Kogan DL, Burroughs M, Emre S, et al: Prospective longitudinal analysis of quantitative Epstein-Barr virus polymerase chain reaction in pediatric liver transplant recipients. Transplantation 1999;67:1068-1070; Green M, Cacciarelli TV, Mazariegos GV, et al: Serial measurement of Epstein-Barr viral load in peripheral blood in lymphoproliferative disease. Transplantation 1998;66:1641-1644; Espy MJ, Patel R, Paya C, Smith TF: Quantification of Epstein-Barr virus viral load in transplant patients by LightCycler PCR. ASM Press 101st General Meeting [Abstract]. 2001;May 20-24)