title: Multicenter evaluation of prototype real-time PCR assays for Epstein-Barr virus and cytomegalovirus DNA in whole blood samples from transplant recipients.
abstract: Quantitative PCR is becoming widespread for diagnosing and monitoring post-transplantation diseases associated with EBV and CMV. These assays need to be standardized to manage patients in different facilities. Five independent laboratories in Japan compared home-brew assays and a prototype assay system to establish a standard quantitative procedure for measuring EBV and CMV. Reference standards and a total of 816 (642 EBV and 174 CMV) whole blood samples from post-transplantation recipients were used for this multicenter evaluation. The prototype reference standard for EBV was compared to a panel of samples, with a theoretical expected value made using EBV-positive cells containing two virus genome copies per cell. The mean ratio of the reference standard at each site to the standard of the prototype assay was ≤ 4.15 for EBV among three different sites and ≤ 3.0 for CMV between two laboratories. The mean of the theoretical expected number of the EBV genome: prototype reference was close to 1.0. The correlation coefficients between the viral copy numbers determined using the prototype assay and those using each home-brew assay were high (EBV, 0.73-0.83, median = 0.78; CMV, 0.54-0.60, median = 0.57). The dynamics of the EBV and CMV loads in transplant recipients were similar between the assay types. There was an inter-laboratory difference among the quantification results, indicating that a unified protocol and kit are favorable for standardizing the quantification of EBV and CMV. Such standardization will help to standardize the diagnosis and monitoring of diseases associated with EBV and CMV.
authors.affiliation: Pediatrics Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan. yoshi-i@med.nagoya-u.ac.jp
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ito
authors.forename: Shunji
authors.initials: S
authors.lastname: Takakura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ichiyama
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Ueda
authors.forename: Yukio
authors.initials: Y
authors.lastname: Ando
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsuda
authors.forename: Eiko
authors.initials: E
authors.lastname: Hidaka
authors.forename: Kaname
authors.initials: K
authors.lastname: Nakatani
authors.forename: Junji
authors.initials: J
authors.lastname: Nishioka
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nobori
authors.forename: Naoki
authors.initials: N
authors.lastname: Kajiyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000097:blood*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007753:Laboratories; D016377:Organ Transplantation; D016133:Polymerase Chain Reaction / Q000592:standards*; D011933:Reagent Kits, Diagnostic; D012015:Reference Standards; D019562:Viral Load
chemical_list: D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic