Quantitation of Hepatitis B Virus Genomic DNA by Real-Time Detection PCR-Reference-Cited by-同舟云学术

Quantitation of Hepatitis B Virus Genomic DNA by Real-Time Detection PCR

Published:1999-09 Issue:9 Volume:37 Page:2899-2903
ISSN:0095-1137
Container-title:Journal of Clinical Microbiology
language:en
Short-container-title:J Clin Microbiol

Author:

Abe Aki¹,Inoue Kazuaki²³,Tanaka Takeshi⁴,Kato Junko²⁵,Kajiyama Naoki¹,Kawaguchi Ryuji¹,Tanaka Satoshi⁴,Yoshiba Makoto³,Kohara Michinori²

Affiliation:

1. Center for Molecular Biology and Cyotogenesis, SRL, Inc., Hino Tokyo 192-0002,1

2. Department of Microbiology, The Tokyo Metropolitan Institute of Medical Science,2 and

3. Division of Gastroenterology, Showa University Fujigaoka Hospital, Aoba-ku, Yokohama 227-8501,3 and

4. Liver Unit, The Tokyo Metropolitan Komagome Hospital,4Bunkyo-ku, Tokyo 113-8613,

5. Institute of Gastroenterology, Tokyo Women’s Medical College, Shinjiyuku-ku, Tokyo 162-0054,5 Japan

Abstract

ABSTRACT Quantitation of hepatitis B virus (HBV) DNA in serum is a useful method for the monitoring of HBV replication. We attempted to develop a quantitative assay system for HBV DNA that is more sensitive, accurate, and reproducible than existing systems. We detected HBV DNA by real-time detection PCR (RTD-PCR) based on Taq Man chemistry. The efficacy of this assay was evaluated by quantitatively measuring sequential levels of synthetic DNA and DNA in clinical serum samples. The detection limit of this system was as few as 10 DNA copies/reaction. A linear standard curve was obtained between 10 1 and 10 8 DNA copies/reaction. The coefficient of variation for both intra- and interexperimental variability indicated remarkable reproducibility. This system detected HBV DNA in 100% of chronic hepatitis B patients tested and never detected HBV DNA in healthy volunteers who were negative for HBV markers. These observations suggest that RTD-PCR is an excellent candidate for a standard HBV quantification method.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

Link

https://journals.asm.org/doi/pdf/10.1128/JCM.37.9.2899-2903.1999

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