QUANTIFICATION OF CYTOMEGALOVIRUS DNA IN BLOOD SPECIMENS FROM BONE-MARROW TRANSPLANT RECIPIENTS BY THE POLYMERASE CHAIN-REACTION

A nested PCR system for cytomegalovirus (CMV) DNA in blood specimens f rom bone marrow transplant recipients is described, in which the bioti nylated tritium-labelled product from the second round of PCR is quant ified using streptavidin-coated fluorometric Scintillation Proximity A ssay (SPA) beads (Amersham, UK). This assay has been compared with a P CR procedure based on limiting-dilution, in which the end-point is det ermined visually following electrophoresis in agarose gel. The two sys tems were shown to be equivalent in sensitivity and specificity on tes ting stored serial blood samples from six CMV antibody-positive alloge neic bone marrow transplant patients who developed viraemia as detecte d by conventional methods of virus isolation in tissue culture.