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Journal of Clinical Microbiology, February 1999, p. 310-314, Vol. 37, No. 2
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Quantitative Detection of Hepatitis B Virus by Transcription-Mediated Amplification and Hybridization Protection Assay

Keiichi Kamisango,1,* Chieko Kamogawa,1 Mayumi Sumi,1 Susumu Goto,1 Akihide Hirao,1 Frank Gonzales,2 Kiyomi Yasuda,3,4 and Shiro Iino3

Diagnostics Research Laboratories, Chugai Diagnostics Science Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171,1 Department of Internal Medicine (4), St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki-shi, Kanagawa 216,3 and Research Center for Liver Diseases, Kiyokawa Hospital, 2-31-12 Asagaya Minami, Suginami-ku, Tokyo 166,4 Japan, and Gen-Probe, Inc., San Diego, California 921212

Received 24 June 1998/Returned for modification 20 August 1998/Accepted 2 November 1998

We have developed a sensitive and quantitative assay using transcription-mediated amplification and hybridization protection assay for the detection of hepatitis B virus (HBV) DNA in serum. The transcription-mediated amplification was carried out in a single tube. The hybridization protection assay was carried out in a microtiter plate with two probes with different specific activities to obtain a broad detection range. As a result, the assay had a detection range of 5 × 103 to 5 × 108 genome equivalents (GE)/ml and good quantitative accuracy on a logarithmic scale. A moderately sized manual assay run can be completed within 5 h. Measurements of the amounts of HBV DNA in clinical samples by the assay showed the amounts under various disease conditions to be widely distributed (more than 5 logs, from approximately 5 × 103 to 5 × 108 GE/ml). It was also shown that the amount of HBV DNA in one chronic hepatitis patient varied widely, with a range of more than 5 logs during long-term monitoring. Our assay has the potential to be used to monitor and determine the prognosis of HBV patients and carriers, especially during interferon treatment.

* Corresponding author. Mailing address: Diagnostics Research Labs., Chugai Diagnostics Science Co. Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171, Japan. Phone: 3-3987-0705. Fax: 3-3989-0785. E-mail: kamisangokii{at}chugai-pharm.co.jp.

Journal of Clinical Microbiology, February 1999, p. 310-314, Vol. 37, No. 2
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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