ArticleAntimicrobial agents and chemotherapy2008
Rolling circle amplification, a powerful tool for genetic and functional studies of complete hepatitis B virus genomes from low-level infections and for directly probing covalently closed circular DNA.
Article in Antimicrobial agents and chemotherapy, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 61 citations in OpenAlex.
- Article
- Primary Duck Hepatocyte Culture and Duck Hepatitis B Virus Infection Model.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Recombinase amplified CRISPR enhanced chain reaction (RACECAR) for viral genome detection.Nanoscale · 2022Article
- Role of hepatitis B virus in development of hepatocellular carcinoma: Focus on covalently closed circular DNA.World journal of hepatology · 2022Review
- Single-stranded DNA aptamer-based rolling circle amplification as anti-chickenVeterinary world · 2022Article
- Rolling Circle Amplification in Integrated Microsystems: An Uncut Gem toward Massively Multiplexed Pathogen Diagnostics and Genotyping.Accounts of chemical research · 2021Review
- A systematic study of Tupaia as a model for human acute hepatitis B infection.The Journal of veterinary medical science · 2021Article
- Quantification and epigenetic evaluation of the residual pool of hepatitis B covalently closed circular DNA in long-term nucleoside analogue-treated patients.Scientific reports · 2020Article
- Amplification chemistries in clinical virology.Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology · 2019Review
- Illumina and Nanopore methods for whole genome sequencing of hepatitis B virus (HBV).Scientific reports · 2019Article
- Quantification of Hepatitis B Virus Covalently Closed Circular DNA in Infected Cell Culture Models by Quantitative PCR.Bio-protocol · 2019Article
- Article
- Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus.PLoS pathogens · 2018Article
- Differences in sequences between HBV-relaxed circular DNA and covalently closed circular DNA.Emerging microbes & infections · 2017Article
- Detection of HBV Covalently Closed Circular DNA.Viruses · 2017Review
- A new model mimicking persistent HBV e antigen-negative infection using covalently closed circular DNA in immunocompetent mice.PloS one · 2017Article
- PCR mediated recombination impacts the analysis of hepatitis B Virus covalently closed circular DNA.Retrovirology · 2016Article
- Article
- Rapid and quantitative detection of hepatitis B virus.World journal of gastroenterology · 2015Review
- A novel method for detection of HBVcccDNA in hepatocytes using rolling circle amplification combined with in situ PCR.BMC infectious diseases · 2014Article
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Complete characterization of the biological properties of hepatitis B virus (HBV) variants requires the generation of full-length genomes. The aim of this study was to develop new tools for the efficient full-length genome amplification of virus from samples with low viral loads. Rolling circle amplification (RCA) was used to amplify full-length HBV genomes from both sera and liver biopsy samples from chronic HBV carriers. Serum-derived relaxed circular HBV DNA could be amplified only after completion and ligation of plus-strand DNA. Covalently closed circular DNA (cccDNA) from liver biopsies could be amplified directly from as few as 13 copies, using RCA, followed by a full-length HBV PCR. Three serial liver biopsy samples were obtained from a lamivudine-resistant patient who cleared detectable serum HBV after adefovir dipivoxil was added to the lamivudine therapy and then seroconverted to anti-HBs. Only the genomes from the last biopsy specimen obtained after the emergence of lamivudine resistance contained the lamivudine resistance-associated mutations rtL180M and rtM204V ("rt" indicates reverse transcriptase domain). Defective genomes were also found in this biopsy sample. Genomes cloned from the liver biopsy specimens were transfected into HuH7 cells to study their replication competence and their susceptibility to lamivudine. RCA is a powerful tool for amplifying full-length HBV genomes and will be especially useful for the study of occult or inactive HBV infections and patients undergoing antiviral treatment. It can also be used to probe HBV cccDNA, the crucial intermediate in viral persistence and the archive of resistance mutations.
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