Korean J Lab Med.
2002 Oct;22(5):325-330.
Hepatitis B Virus DNA Assay using the TaqMan PCR System
- Affiliations
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- 1Department of Virology, University Hospital Rotterdam, Rotterdam, The Netherlands.
- 2Department of Laboratory Medicine and Institute of Wonkwang Medical Science, School of Medicine, Wonkwang University, Iksan, Korea. email@wmc.wonkwang.ac.kr
- 3Department of Virology, Helsinki University, Central Hospital, Helsinki, Finland.
- 4Veterinary Internal Medicine/Clinical Pathology, College of Veterinary Medicine, Chonbuk National University, Chonju, Korea.
Abstract
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BACKGROUND: Hepatitis B virus (HBV) DNA quantitation and detection assay is necessary when monitoring the disease progression or therapeutic effect of viral hepatitis. The TaqMan PCR system (TPS) that was recently developed for HBV DNA quantitation was known for not only capable and exact quantitation but also capable sensitive detection. Therefore, we evaluated the clinical utility of the TPS by comparing it with hybrid capture system (HCS) and nested polymerase chain reaction (PCR).
METHODS
The dilution test was performed to evaluate reproducibility and the dynamic range of TPS and HCS. Further, the sensitivity of the diluted sera were also compared with TPS, HCS, and nested PCR. The sera of HBsAg positive patients (n=119) were tested to compare the sensitivity of the three methods and the quantity of HBV DNA by TPS and HCS.
RESULTS
The TPS showed lower coefficients of variations (TPS, 6.5- 60.4%; HCS, 11.9-61.4%) and a wider dynamic range than HCS in the dilution test. The sensitivity was high in order of nested PCR, TPS and HCS (cover 10(6), 5 X 10(5), and 10(2) of diluted concentration) in the dilution test. But the sensitivity was high in order of TPS, nested PCR, and HCS (89, 68%, and 56%) in the 119 sera of HBsAg positive patients. The TPS and HCS revealed a significant correlation (R=0.90, P<0.0001).
CONCLUSIONS
The TPS and nested PCR showed higher sensitivity than HCS, and TPS showed better sensitivity than nested PCR in clinical specimens. Also, the TPS showed more quantitation potential than HCS. Thus, it appears useful in accurately defining the viral replication state and antiviral therapeutic effects among persons with HBV infection.
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