ArticleBiochemical and biophysical research communications2006
Efficient amplification and cloning of near full-length hepatitis C virus genome from clinical samples.
Article in Biochemical and biophysical research communications, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- A method for near full-length amplification and sequencing for six hepatitis C virus genotypes.BMC genomics · 2016Article
- Article
- Comprehensive cloning of patient-derived 9022-bp amplicons of hepatitis C virus.Journal of virological methods · 2013Article
- Viral categorization and discovery in human circulation by transcriptome sequencing.Biochemical and biophysical research communications · 2013Article
- Development of a sensitive RT-PCR method for amplifying and sequencing near full-length HCV genotype 1 RNA from patient samples.Virology journal · 2013Article
- De novo polymerase activity and oligomerization of hepatitis C virus RNA-dependent RNA-polymerases from genotypes 1 to 5.PloS one · 2011Article
- Enhanced protocol for determining the 3' terminus of hepatitis C virus.Journal of virological methods · 2010Article
- Comparative analysis of nearly full-length hepatitis C virus quasispecies from patients experiencing viral breakthrough during antiviral therapy: clustered mutations in three functional genes, E2, NS2, and NS5a.Journal of virology · 2008Article
- Hepatitis C virus diversity and evolution in the full open-reading frame during antiviral therapy.PloS one · 2008Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Long RT-PCR (LRP) amplification of RNA templates is sometimes difficult compared to long PCR of DNA templates. Among RNA templates, hepatitis C virus (HCV) represents an excellent example to challenge the potential of LRP technology due to its extensive secondary structures and its difficulty to be readily cultured in vitro. The only source for viral genome amplification is clinical samples in which HCV is usually present at low titers. We have created a comprehensive optimization protocol that allows robust amplification of a 9.1 kb fragment of HCV, followed by efficient cloning into a novel vector. Detailed analyses indicate the lack of potential LRP-mediated recombination and the preservation of viral diversity. Thus, our LRP protocol could be applied for the amplification of other difficult RNA templates and may facilitate RNA virus research such as linked viral mutations and reverse genetics.
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Registered trials
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