HBV DNA Quantitation Using Real-time PCR.
10.3343/kjlm.2006.26.6.424
- Author:
Jeong HEO
1
;
Won Ook GO
;
Gwang Ha KIM
;
Dae Hwan KANG
;
Geun Am SONG
;
Mong CHO
;
Hyung Hoi KIM
;
Eeu Yup LEE
Author Information
1. Department of Internal Medicine, Pusan National University, School of Medicine, Busan, Korea.
- Publication Type:Original Article
- Keywords:
HBV DNA quantitation;
Real time PCR;
Cobas amplicor
- MeSH:
Calibration;
DNA*;
Hepatitis B virus;
Humans;
Real-Time Polymerase Chain Reaction*
- From:The Korean Journal of Laboratory Medicine
2006;26(6):424-430
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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 x 10(4) 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 Monitor(TM) 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 x 10(2) copies/mL for HBV, and the quantitation was accurate and reproducible over a wide dynamic range from 6.1 x 0(2) to 6.5 x 10(9) 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 Monitor(TM) kit. CONCLUSIONS: 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.
