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2030-01-01
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- IOC - Artigos de Periódicos [12488]
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IMPROVED ROLLING CIRCLE AMPLIFICATION (RCA) OF HEPATITIS B VIRUS (HBV) RELAXED-CIRCULAR SERUM DNA (RC-DNA)
Serum relaxed-circular DNA
Rolling-Circle Amplification (RCA)
HBV genotypes
Recombinant genomes
Affilliation
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France / Hôpital de la Croix Rousse. Hepato-Gastroenterology Unit. Lion, France.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France / Hôpital de la Croix Rousse. Hepato-Gastroenterology Unit. Lion, France.
Centre de Recherche en Cancérologie de Lyon (CRCL). INSERM, U1052; CNRS, UMR 5286; UCBL1, S 1052. 151 cours Albert Thomas, 69003 Lyon, France.
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 >1.0E+8 IU/ml down to 1.0E+3 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
HBVSerum relaxed-circular DNA
Rolling-Circle Amplification (RCA)
HBV genotypes
Recombinant genomes
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