Brasil | Article

A new method for next-generation sequencing of the full Hepatitis B virus genome from a clinical specimen: impact for vitus genotyping

dc.creatorBarbosa, Flavia Hebeler
dc.creatorWolf, Ivan Rodrigo
dc.creatorValente, Guilherme Targino
dc.creatorMello, Francisco Campello do Amaral
dc.creatorLampe, Elisabeth
dc.creatorPardini, Maria Inês de Moura Campos
dc.creatorGrotto, Rejane Maria Tommasini
dc.date2020-12-28T14:54:22Z
dc.date2020-12-28T14:54:22Z
dc.date2020
dc.date.accessioned2023-09-27T00:10:53Z
dc.date.available2023-09-27T00:10:53Z
dc.identifierBARBOSA, Flavia Hebeler et al. A new method for next-generation sequencing of the full Hepatiti B virus genome from a clinical specimen: impact for virus genotyping. Microorganims, v. 8, n.1291, p. 1-12, Sept. 2020.
dc.identifier2076-2607
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/45445
dc.identifier10.3390/microorganisms8091391
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8898254
dc.descriptionHepatitis B virus (HBV) is an enveloped virus that induces chronic liver disease. HBV has been classified into eight genotypes (A–H) according to its genome sequence by using Sanger sequencing or reverse hybridization. Sanger sequencing is often restricted to analyzing the S gene and is inaccurate for detecting minority genetic variants, whereas reverse hybridization detects only known mutations. Next-generation sequencing (NGS) is a robust tool for clinical virology with different protocols available. The objective of this study was to develop a new method for the study of viral genetic polymorphisms or more accurate genotyping using genome amplification followed by NGS. Plasma obtained from five chronically infected HBV individuals was used for viral DNA isolation. HBV full-genome PCR amplification was the enrichment method for NGS. Primers were used to amplify all HBV genotypes in three overlapping amplicons, following a tagmentation step and Illumina NGS. For phylogenetic analysis, sequences were extracted from the HBVdb database. We were able to amplify a full HBV genome; further, NGS was shown to be a robust method and allowed better genotyping, mainly in patients carrying mixed genotypes, classified according to other techniques. This new method may be significant for whole genome analyses, including other viruses.
dc.formatapplication/pdf
dc.languageeng
dc.publisherMDPI
dc.rightsopen access
dc.subjectVírus da Hepatite B
dc.subjectGenotipagem
dc.subjectNGS
dc.subjectAnálise filogenia
dc.subjectHepatitis B virus
dc.subjectNGS
dc.subjectGenotyping
dc.subjectPhylogeny analysis
dc.titleA new method for next-generation sequencing of the full Hepatitis B virus genome from a clinical specimen: impact for vitus genotyping
dc.typeArticle


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