| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 6134124 | Journal of Virological Methods | 2013 | 7 Pages |
Abstract
For functional analysis of HBV isolates, epidemiological studies and correct identification of recombinant genomes, the amplification of complete genomes is necessary. A method for completely in vitro amplification of full-length HBV genomes starting from serum RC-DNA is described. This uses in vitro completion/ligation of plus-strand HBV RC-DNA and amplification using Rolling-Circle Amplification, eventually followed by a genomic PCR. The method can amplify complete HBV genomes from sera with viral loads ranging from >1.0EÂ +Â 8Â IU/ml down to 1.0EÂ +Â 3Â IU/ml. The method can be applied to archived sera that have undergone long-term storage or to archived DNA serum extracts. The genomes can easily be cloned. HBV genotypes A-G can all be amplified with no apparent problems. A recombinant subgenotype A3/genotype E genome was identified and fully sequenced.
Keywords
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Authors
Nora Martel, Selma A. Gomes, Isabelle Chemin, Christian Trépo, Alan Kay,