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KMID : 0981820060260060424
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Korean Journal of Laboratory Medicine
2006 Volume.26 No. 6 p.424 ~ p.430
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HBV DNA Quantitation Using Real-time PCR
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Heo Jeong
Go Won-Ook
Kim Gwang-Ha
Kang Dae-Hwan
Song Geun-Am
Cho Mong
Kim Hyung-Hoi
Lee Eun-Yup
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Abstract
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Background: Accurate measurement of the concentration of hepatitis B virus (HBV) DNA in clinical samples is important for the appropriate treatment of patients and evaluation of their therapeutic responses. In addition, the concentration of HBV DNA in the serum of patients with chronic HBV infection has a very broad range. Real-time PCR is very sensitive and useful to detect HBV DNA in a wide range of concentrations. We designed new primers and probes for real-time PCR to detect HBV DNA.
Methods: Primers and probes specific for HBV were designed. EUROHEP HBV DNA standards (NIBSC, Hertfordshire, UK) with the HBV DNA concentration of 7.0×104 copies/mL was used to determine the calibration curve and efficacy for the real-time PCR assay. Sensitivity, dynamic range, and precision were evaluated. The correlation between the real-time PCR and Cobas Amplicor HBV MonitorTM assay in the measurement of serum HBV DNA concentrations in 52 patients with chronic HBV infection was evaluated.
Results: The sensitivity of the assay was approximately 6.08×102 copies/mL for HBV, and the quantitation was accurate and reproducible over a wide dynamic range from 6.1×02 to 6.5×109 copies/mL without any dilution of specimens. The assay showed low coefficients of variation of repeatability (3.7-24.9%) and reproducibility (7.8-24.7%). The results were found to correlate well with those obtained by Cobas Amplicor HBV MonitorTM kit.
Conclusion: We provide a new in-house method for the measurement of serum HBV DNA using real-time PCR, which enables us to detect HBV DNA rapidly, sensitively, and accurately. (Korean J Lab Med 2006;26:424-30)
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KEYWORD
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HBV DNA quantitation, Real time PCR, Cobas amplicor
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