Development and Evaluation of a Real-Time TaqMan-PCR for the Detection of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients.

Cytomegalovirus (CMV) has been recognized as the most important viral pathogen in persons undergoing bone marrow transplantation (BMT). In this study, we present the development of a TaqMan-based real-time PCR assay to quantify human cytomegalovirus (CMV) DNA in peripheral blood leukocytes (PBLs) of bone marrow transplantation patients. A plasmid containing the target sequence from the pp65 region (UL83) of CMV was constructed as a positive control template. Serial dilutions of 107 to 101 plasmids per assay were prepared. Peripheral blood samples were collected from patients after transplantation. CMV DNA was quantified by RQ-PCR in parallel with the pp65 antigenemia assay in PBL samples. The real-time PCR assay could detect CMV DNA in patient''s samples with a wide linear range, from 10 to over 107 copies of CMV. Real-time PCR assay results correlated with those of the CMV pp65 antigenemia assay (P < 0.0001). The TaqMan assay may be a useful tool for rapid quantification of CMV infection and for monitoring of CMV reactivation in bone marrow transplantation recipients. The results of both quantitative assays were significantly correlated; however, the RQ-PCR assay was more sensitive than the pp65 antigenemia assay.