Development of a real-time quantitative assay for detection of Epstein-Barr virus

With the use of real-time PCR, we developed and evaluated a rapid, sensitiv e, specific, and reproducible method for the detection of Epstein-Barr viru s (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and 10(7) copies of DNA per mi using two sample preparation methods based on absorption. ri precision study yie lded an average coefficient of variation for both methods of less than 12%, with a coefficient of regression for the standard curve of a minimum of 0. 98. We detected EBV DNA in 19.2% of plasma samples from immunosuppressed so lid-organ transplant patients without symptoms of EBV infections with a mea n load of 440 copies per mi, EBV DNA could be detected in all transplant pa tients diagnosed with posttransplant lymphoproliferative disorder, with a m ean load of 544,570 copies per mi, No EBV DNA could be detected in healthy individuals in nonimmunosuppressed control groups and a mean of 6, 100 copi es per mi could be detected in patients with infectious mononucleosis. Furt her studies revealed that the inhibitory effect of heparinized plasma could be efficiently removed by use of an extraction method with Celite as the a bsorbent.