A Genotype-Independent Real-Time PCR Assay for Quantification of Hepatitis B Virus DNA
- 1 February 2007
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 45 (2), 553-558
- https://doi.org/10.1128/jcm.00709-06
Abstract
Accurate quantification of hepatitis B virus (HBV) DNA levels is important for monitoring patients with chronic HBV infection and for assessing their responses to antiviral therapy. This study aimed to develop a real-time PCR assay that is sensitive and can accurately quantify a wide range of HBV DNA levels across the known HBV genotypes. An “in-house” real-time PCR assay using primers and a TaqMan probe in a highly conserved region of the HBV surface gene was designed. The assay was standardized against a WHO standard and validated against plasmids of HBV genotypes A through H. The linear quantification range was approximately 5 × 10 0 to 2.0 × 10 9 IU/ml. Results of samples from patients infected with HBV genotypes A through H tested using our real-time “in-house” PCR assay showed an excellent correlation with those of the Cobas Amplicor HBV Monitor ( R 2 = 0.9435) and the Cobas TaqMan HBV ( R 2 = 0.9873) tests. We have established a real-time PCR assay that is genotype independent and can accurately quantify a wide range of HBV DNA levels. Further studies of additional samples are ongoing to validate the genotype independence of our assay.Keywords
This publication has 25 references indexed in Scilit:
- Comparison of the COBAS TaqMan™ HBV test with the COBAS Amplicor monitor™ test for measurement of hepatitis B virus DNA in serumJournal of Medical Virology, 2005
- A 1-Year Trial of Telbivudine, Lamivudine, and the Combination in Patients With Hepatitis B e Antigen—Positive Chronic Hepatitis BGastroenterology, 2005
- Quantification and genotyping of hepatitis B virus in a single reaction by real-time PCR and melting curve analysisJournal of Hepatology, 2004
- Hepatitis B virus genotypes: Do they play a role in the outcome of HBV infection?Hepatology, 2004
- Hepatitis B virus genotypes: Do they play a role in the outcome of HBV infection?Hepatology, 2004
- Quantitative serum HBV DNA levels during different stages of chronic hepatitis B infectionHepatology, 2002
- Quantitative serum HBV DNA levels during different stages of chronic hepatitis B infectionHepatology, 2002
- Quantitative Analysis of Hepatitis B Virus DNA by Real-Time AmplificationEuropean Journal of Clinical Microbiology & Infectious Diseases, 2001
- A comparison of reverse transcription–polymerase chain reaction and branched‐chain DNA assays for hepatitis C virus RNA in patients receiving interferon treatmentJournal of Viral Hepatitis, 1999
- The ?gold-standard,? accuracy, and the current concepts: Hepatitis C virus genotype and viremiaHepatology, 1996