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Journal of Clinical Microbiology, July 2000, p. 2536-2542, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Real-Time Automated PCR for Early Diagnosis and Monitoring of Cytomegalovirus Infection after Bone Marrow Transplantation

Utako Machida,1 Masahiro Kami,1,2 Takafumi Fukui,3 Yukumasa Kazuyama,3 Moritoshi Kinoshita,3 Yuji Tanaka,1 Yoshinobu Kanda,1 Seishi Ogawa,1 Hiroaki Honda,1 Shigeru Chiba,1 Kinuko Mitani,1 Yoshitomo Muto,2 Kazuoki Osumi,3 Satoshi Kimura,4 and Hisamaru Hirai1,*

Department of Hematology and Oncology1 and Department of Infectious Diseases,4 Faculty of Medicine, The University of Tokyo; Department of Hematology, Toranomon Hospital2; and Otsuka Assay Laboratories, Otsuka Pharmaceutical Co., Ltd.,3 Tokyo, Japan

Received 30 August 1999/Returned for modification 18 November 1999/Accepted 1 May 2000

The purpose of this study was to assess the usefulness of real-time automated PCR as a quantitative, highly reproducible, and sensitive method to detect cytomegalovirus (CMV) DNA in blood specimens. Intra- and interassay precision rates were 0.89% (small number of copies [L]), 1.43% (middle number of copies [M]), and 1.12% (high number of copies [H]), and 4.46% (L), 1.51% (M), and 2.28% (H), respectively. The linearity of this assay was obtained between 10 and 107 copies/well, with a minimum detection limit of 20 copies/well. Specimens from 55 of 70 healthy subjects were found to be positive for CMV antibody, but CMV DNA was not detected in any of them. In the qualitative assessment of each specimen, the results of the CMV antigenemia assay and those of the real-time PCR assay agreed in 80% (plasma specimens), 79% (all nucleated cells), and 86% (blood) of the cases examined. For eight patients diagnosed as having CMV infection or disease, no sample was positive in the antigenemia assay earlier than in the real-time PCR assay. Furthermore, the results of this assay could be obtained within 8 h. We concluded that the real-time PCR assay is useful for rapid diagnosis of CMV infection and monitoring of clinical courses.

* Corresponding author. Mailing address: Department of Hematology and Oncology, Faculty of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8655, Japan. Phone: 81-3-3815-5411, ext. 5600. Fax: 81-3-5680-7286. E-mail: hhirai-tky{at}umin.ac.jp.

Journal of Clinical Microbiology, July 2000, p. 2536-2542, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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