Quantification of Human Cytomegalovirus DNA by a New Capture Hybrid Polymerase Chain Reaction Enzyme-Linked Immunosorbent Assay in Plasma and Peripheral Blood Mononuclear Cells of Bone Marrow Transplant Recipients

Abstract

Objectives: Quantitative monitoring of human cytomegalovirus infections is helpful in determining appropriate antiviral management in patients who receive bone marrow transplants. We sought to design and evaluate a new cytomegalovirus capture hybrid polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) in plasma and peripheral blood mononuclear cells to monitor cytomegalovirus infection in bone marrow transplant recipients.

Patients and Methods: Twenty-six patients who received allogeneic bone marrow transplants, including 17 male patients and 9 female patients (9 adults, 17 children), were enrolled in this study. A total of 313 consecutive whole blood specimens, before and from 7 to 120 days after transplant, was evaluated in the study. A newly designed biotinylated probe-mediated quantitative competitive PCR-ELISA test was used to determine cyto­megalovirus load in specimens of peripheral blood mononuclear cells and plasma.

Results: All 26 patients were cytomegalovirus seropositive before transplant. Capture hybrid PCR-ELISA of peripheral blood mononuclear cells detected cytomegalovirus DNA in 287 of 313 specimens (91.7%) even in cases with no active cytomegalovirus infection. in plasma, cyto­megalovirus DNA was detected in 114 of 313 specimens (36.4%). Increasing titers of cyto­megalovirus DNA were detected in 14 of 26 patients (53.8%).

Conclusions: The quantitative capture hybrid PCR-ELISA was able to diagnose and monitor cytomegalovirus infection in patients who received bone marrow transplants. Detection of cyto­megalovirus DNA in plasma was more predictive of the onset of cytomegalovirus-related clinical symptoms, compared to detection in peripheral blood mononuclear cells.