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Journal of Clinical Microbiology, January 1999, p. 132-136, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

Hiroshi Kimura,1,* Makoto Morita,1 Yumi Yabuta,1 Kiyotaka Kuzushima,2 Koji Kato,3 Seiji Kojima,3 Takaharu Matsuyama,3 and Tsuneo Morishima1

Department of Pediatrics, Nagoya University School of Medicine,1 Laboratory of Viral Oncology, Research Institute, Aichi Cancer Center,2 and Division of Hematology/Oncology, Children's Medical Center, Japanese Red Cross Nagoya First Hospital,3 Nagoya, Japan

Received 2 July 1998/Returned for modification 20 August 1998/Accepted 13 October 1998

To measure the virus load in patients with symptomatic Epstein-Barr virus (EBV) infections, we used a real-time PCR assay to quantify the amount of EBV DNA in blood. The real-time PCR assay could detect from 2 to over 107 copies of EBV DNA with a wide linear range. We estimated the virus load in peripheral blood mononuclear cells (PBMNC) from patients with symptomatic EBV infections. The mean EBV-DNA copy number in the PBMNC was 103.7 copies/µg of DNA in patients with EBV-related lymphoproliferative disorders, 104.1 copies/µg of DNA in patients with chronic active EBV infections, and 102.2 copies/µg of DNA in patients with infectious mononucleosis. These numbers were significantly larger than those in either posttransplant patients or immunocompetent control patients without EBV-related diseases. In a patient with infectious mononucleosis, the virus load decreased as the symptoms resolved. The copy number of EBV DNA in PBMNC from symptomatic EBV infections was correlated with the EBV-positive cell number determined by the in situ hybridization assay (r = 0.842; P < 0.0001). These results indicate that the real-time PCR assay is useful for diagnosing symptomatic EBV infection and for monitoring the virus load.

* Corresponding author. Mailing address: Department of Pediatrics, Nagoya University School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. Phone: 81-52-744-2303. Fax: 81-52-744-2974. E-mail: hkimura{at}med.nagoya-u.ac.jp.

Journal of Clinical Microbiology, January 1999, p. 132-136, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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