title: Construction and Evaluation of Cytomegalovirus DNA Quantification System with Real-Time Detection Polymerase Chain Reaction.
abstract: For patients with reactivation of human cytomegalovirus (CMV), a highly sensitive and accurate CMV quantification system is essential to monitor viral load.
  
  
  
  We constructed a real-time detection PCR (RTD-PCR) system for CMV DNA and evaluated its linearity, lower detection limit, dynamic range and accuracy using two CMV standards. We used 219 clinical samples derived from 101 patients to compare the system with the pp65 antigen test.
  
  
  
  The 95% detection limit was determined to be 556 IU/mL (95% CI, 440-797 IU/mL), and the quantification range was between 102and 106copies or IU/mL (r= 0.996, 0.999, respectively). The coefficients of variation of inter-assay reproducibility assessed in each three different runs were 2.5% at 1,000 IU/mL and 1.6% at 10,000 IU/mL. The coefficients of variation of intra-assay variability by testing the same samples three times in a single run were 1.8-3.6% and 0.4-1.9%, respectively. The concordance between antigenemia and plasma or serum CMV DNA levels was a good correlation (r= 0.695,P< 0.01).
  
  
  
  We constructed the RTD-PCR system which enables accurate evaluation of CMV reactivation by monitoring of viral load in immunosuppressed or immunocompromised patients.
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hatayama
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hashimoto
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Hara
authors.affiliation: Division of Clinical Laboratory, Tottori University Hospital, Yonago 683-8504, Japan; †Division of Clinical Laboratory Medicine, Department of Pathophysiological and Therapeutic Science, School of Medicine, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Motokura
keywords: CMV antigenemia; RTD-PCR; cytomegalovirus; quantification