Multicenter evaluation of prototype real‐time PCR assays for Epstein‐Barr virus and cytomegalovirus DNA in whole blood samples from transplant recipients |
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Authors: | Yoshinori Ito Shunji Takakura Satoshi Ichiyama Mitsuharu Ueda Yukio Ando Kazuyuki Matsuda Eiko Hidaka Kaname Nakatani Junji Nishioka Tsutomu Nobori Naoki Kajiyama Hiroshi Kimura |
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Institution: | 1. Pediatrics;2. Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, Kyoto 606‐8501;3. Department of Diagnostic Medicine, Kumamoto University Graduate School of Medicine, Kumamoto 860‐8556;4. Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto 390‐8621;5. Department of Molecular and Laboratory Medicine, Mie University Graduate School of Medicine, Tsu 514‐8507;6. Roche Diagnostics K.K., Tokyo 105‐0014, Japan;7. Virology, Nagoya University Graduate School of Medicine, Nagoya 466‐8550 |
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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. |
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Keywords: | cytomegalovirus Epstein– Barr virus real‐time PCR standardization viral load |
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