A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus
Resumen:
Torque Teno Virus (TTV) was initially associated with post-transfusion hepatitis, but growing evidence of its ubiquity in humans is compatible to no apparent clinical significance. TTV is a small non-enveloped virus with a circular single-negative-stranded DNA genome, belonging to the Anelloviridae family. Currently, TTVs are divided in seven phylogenetic groups and are further classified into 21 species. Studies about diversity of TTV in different conditions are receiving increasing interest and in this sense, sequencing of whole genomes for better genetic characterization becomes even more important. Since its discovery in 1997, few TTV complete genomes have been reported worldwide. This is probably due, among other reasons, to the great genetic heterogeneity among TTV strains that prevents its amplification and sequencing by conventional PCR and cloning methods. In addition, although metagenomics approach is useful in these cases, it remains a challenging tool for viromic analysis. With the aim of contributing to the expansion of the TTV whole genomes dataset and to study intra-host variants, we employed a methodology that combined a rolling-circle amplification approach followed by EcoRI digestion, generating a DNA fragment of ~4Kb consistent with TTV genome length which was sequenced by Illumina next generation sequencing. A genogroup 3 full-length consensus TTV genome was obtained and co-infection with other species (at least those with a single EcoRI cleavage site) was not identified. Additionally, bioinformatics analysis allowed to identify the spectrum of TTV intra-host variants which provides evidence of a complex evolution dynamics of these DNA circular viruses, similarly to what occurs with RNA viruses.
2023 | |
Torque Teno Virus Whole genome Rolling circle-amplification Restriction endonuclease digestion Next generation sequencing Intra-host variants |
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Inglés | |
Universidad de la República | |
COLIBRI | |
https://hdl.handle.net/20.500.12008/42749 | |
Acceso abierto | |
Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
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---|---|
author | Cancela D'Angelo, Florencia |
author2 | Marandino, Ana Panzera Crespo, Yanina Bentancour, Gabriela Mirazo, Santiago Arbiza, Juan Ramos, Natalia |
author2_role | author author author author author author |
author_facet | Cancela D'Angelo, Florencia Marandino, Ana Panzera Crespo, Yanina Bentancour, Gabriela Mirazo, Santiago Arbiza, Juan Ramos, Natalia |
author_role | author |
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collection | COLIBRI |
dc.contributor.filiacion.none.fl_str_mv | Cancela D'Angelo Florencia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. Marandino Ana, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. Panzera Crespo Yanina, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. Bentancour Gabriela, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. Mirazo Santiago, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Química Biológica. Arbiza Juan, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. Ramos Natalia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. |
dc.creator.none.fl_str_mv | Cancela D'Angelo, Florencia Marandino, Ana Panzera Crespo, Yanina Bentancour, Gabriela Mirazo, Santiago Arbiza, Juan Ramos, Natalia |
dc.date.accessioned.none.fl_str_mv | 2024-02-27T14:33:57Z |
dc.date.available.none.fl_str_mv | 2024-02-27T14:33:57Z |
dc.date.issued.none.fl_str_mv | 2023 |
dc.description.abstract.none.fl_txt_mv | Torque Teno Virus (TTV) was initially associated with post-transfusion hepatitis, but growing evidence of its ubiquity in humans is compatible to no apparent clinical significance. TTV is a small non-enveloped virus with a circular single-negative-stranded DNA genome, belonging to the Anelloviridae family. Currently, TTVs are divided in seven phylogenetic groups and are further classified into 21 species. Studies about diversity of TTV in different conditions are receiving increasing interest and in this sense, sequencing of whole genomes for better genetic characterization becomes even more important. Since its discovery in 1997, few TTV complete genomes have been reported worldwide. This is probably due, among other reasons, to the great genetic heterogeneity among TTV strains that prevents its amplification and sequencing by conventional PCR and cloning methods. In addition, although metagenomics approach is useful in these cases, it remains a challenging tool for viromic analysis. With the aim of contributing to the expansion of the TTV whole genomes dataset and to study intra-host variants, we employed a methodology that combined a rolling-circle amplification approach followed by EcoRI digestion, generating a DNA fragment of ~4Kb consistent with TTV genome length which was sequenced by Illumina next generation sequencing. A genogroup 3 full-length consensus TTV genome was obtained and co-infection with other species (at least those with a single EcoRI cleavage site) was not identified. Additionally, bioinformatics analysis allowed to identify the spectrum of TTV intra-host variants which provides evidence of a complex evolution dynamics of these DNA circular viruses, similarly to what occurs with RNA viruses. |
dc.format.extent.es.fl_str_mv | 6 h. |
dc.format.mimetype.es.fl_str_mv | application/pdf |
dc.identifier.citation.es.fl_str_mv | Cancela D'Angelo, F, Marandino, A, Panzera Crespo, Y [y otros autores]. "A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus". Virus Research. [en línea] 2023, 323: 198974. 6 h. DOI: 10.1016/j.virusres.2022.198974. |
dc.identifier.doi.none.fl_str_mv | 10.1016/j.virusres.2022.198974 |
dc.identifier.issn.none.fl_str_mv | 0168-1702 |
dc.identifier.uri.none.fl_str_mv | https://hdl.handle.net/20.500.12008/42749 |
dc.language.iso.none.fl_str_mv | en eng |
dc.publisher.es.fl_str_mv | Elsevier |
dc.relation.ispartof.es.fl_str_mv | Virus Research, 2023, 323: 198974. |
dc.rights.license.none.fl_str_mv | Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess |
dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
dc.subject.es.fl_str_mv | Torque Teno Virus Whole genome Rolling circle-amplification Restriction endonuclease digestion Next generation sequencing Intra-host variants |
dc.title.none.fl_str_mv | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
dc.type.es.fl_str_mv | Artículo |
dc.type.none.fl_str_mv | info:eu-repo/semantics/article |
dc.type.version.none.fl_str_mv | info:eu-repo/semantics/publishedVersion |
description | Torque Teno Virus (TTV) was initially associated with post-transfusion hepatitis, but growing evidence of its ubiquity in humans is compatible to no apparent clinical significance. TTV is a small non-enveloped virus with a circular single-negative-stranded DNA genome, belonging to the Anelloviridae family. Currently, TTVs are divided in seven phylogenetic groups and are further classified into 21 species. Studies about diversity of TTV in different conditions are receiving increasing interest and in this sense, sequencing of whole genomes for better genetic characterization becomes even more important. Since its discovery in 1997, few TTV complete genomes have been reported worldwide. This is probably due, among other reasons, to the great genetic heterogeneity among TTV strains that prevents its amplification and sequencing by conventional PCR and cloning methods. In addition, although metagenomics approach is useful in these cases, it remains a challenging tool for viromic analysis. With the aim of contributing to the expansion of the TTV whole genomes dataset and to study intra-host variants, we employed a methodology that combined a rolling-circle amplification approach followed by EcoRI digestion, generating a DNA fragment of ~4Kb consistent with TTV genome length which was sequenced by Illumina next generation sequencing. A genogroup 3 full-length consensus TTV genome was obtained and co-infection with other species (at least those with a single EcoRI cleavage site) was not identified. Additionally, bioinformatics analysis allowed to identify the spectrum of TTV intra-host variants which provides evidence of a complex evolution dynamics of these DNA circular viruses, similarly to what occurs with RNA viruses. |
eu_rights_str_mv | openAccess |
format | article |
id | COLIBRI_9b47bcab4331baf62d6a979cbedfba14 |
identifier_str_mv | Cancela D'Angelo, F, Marandino, A, Panzera Crespo, Y [y otros autores]. "A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus". Virus Research. [en línea] 2023, 323: 198974. 6 h. DOI: 10.1016/j.virusres.2022.198974. 0168-1702 10.1016/j.virusres.2022.198974 |
instacron_str | Universidad de la República |
institution | Universidad de la República |
instname_str | Universidad de la República |
language | eng |
language_invalid_str_mv | en |
network_acronym_str | COLIBRI |
network_name_str | COLIBRI |
oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/42749 |
publishDate | 2023 |
reponame_str | COLIBRI |
repository.mail.fl_str_mv | mabel.seroubian@seciu.edu.uy |
repository.name.fl_str_mv | COLIBRI - Universidad de la República |
repository_id_str | 4771 |
rights_invalid_str_mv | Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
spelling | Cancela D'Angelo Florencia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Marandino Ana, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Panzera Crespo Yanina, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Bentancour Gabriela, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Mirazo Santiago, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Química Biológica.Arbiza Juan, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Ramos Natalia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.2024-02-27T14:33:57Z2024-02-27T14:33:57Z2023Cancela D'Angelo, F, Marandino, A, Panzera Crespo, Y [y otros autores]. "A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus". Virus Research. [en línea] 2023, 323: 198974. 6 h. DOI: 10.1016/j.virusres.2022.198974.0168-1702https://hdl.handle.net/20.500.12008/4274910.1016/j.virusres.2022.198974Torque Teno Virus (TTV) was initially associated with post-transfusion hepatitis, but growing evidence of its ubiquity in humans is compatible to no apparent clinical significance. TTV is a small non-enveloped virus with a circular single-negative-stranded DNA genome, belonging to the Anelloviridae family. Currently, TTVs are divided in seven phylogenetic groups and are further classified into 21 species. Studies about diversity of TTV in different conditions are receiving increasing interest and in this sense, sequencing of whole genomes for better genetic characterization becomes even more important. Since its discovery in 1997, few TTV complete genomes have been reported worldwide. This is probably due, among other reasons, to the great genetic heterogeneity among TTV strains that prevents its amplification and sequencing by conventional PCR and cloning methods. In addition, although metagenomics approach is useful in these cases, it remains a challenging tool for viromic analysis. With the aim of contributing to the expansion of the TTV whole genomes dataset and to study intra-host variants, we employed a methodology that combined a rolling-circle amplification approach followed by EcoRI digestion, generating a DNA fragment of ~4Kb consistent with TTV genome length which was sequenced by Illumina next generation sequencing. A genogroup 3 full-length consensus TTV genome was obtained and co-infection with other species (at least those with a single EcoRI cleavage site) was not identified. Additionally, bioinformatics analysis allowed to identify the spectrum of TTV intra-host variants which provides evidence of a complex evolution dynamics of these DNA circular viruses, similarly to what occurs with RNA viruses.Submitted by Pintos Natalia (nataliapintosmvd@gmail.com) on 2024-02-26T13:57:29Z No. of bitstreams: 2 license_rdf: 25790 bytes, checksum: 489f03e71d39068f329bdec8798bce58 (MD5) 10.1016j.virusres.2022.198974.pdf: 2620713 bytes, checksum: 17604354c6f76b4a2e427d779e5696de (MD5)Approved for entry into archive by Faget Cecilia (lfaget@fcien.edu.uy) on 2024-02-27T13:01:06Z (GMT) No. of bitstreams: 2 license_rdf: 25790 bytes, checksum: 489f03e71d39068f329bdec8798bce58 (MD5) 10.1016j.virusres.2022.198974.pdf: 2620713 bytes, checksum: 17604354c6f76b4a2e427d779e5696de (MD5)Made available in DSpace by Luna Fabiana (fabiana.luna@seciu.edu.uy) on 2024-02-27T14:33:57Z (GMT). No. of bitstreams: 2 license_rdf: 25790 bytes, checksum: 489f03e71d39068f329bdec8798bce58 (MD5) 10.1016j.virusres.2022.198974.pdf: 2620713 bytes, checksum: 17604354c6f76b4a2e427d779e5696de (MD5) Previous issue date: 20236 h.application/pdfenengElsevierVirus Research, 2023, 323: 198974.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)Torque Teno VirusWhole genomeRolling circle-amplificationRestriction endonuclease digestionNext generation sequencingIntra-host variantsA combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virusArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaCancela D'Angelo, FlorenciaMarandino, AnaPanzera Crespo, YaninaBentancour, GabrielaMirazo, SantiagoArbiza, JuanRamos, NataliaLICENSElicense.txtlicense.txttext/plain; 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- Universidad de la Repúblicafalse |
spellingShingle | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus Cancela D'Angelo, Florencia Torque Teno Virus Whole genome Rolling circle-amplification Restriction endonuclease digestion Next generation sequencing Intra-host variants |
status_str | publishedVersion |
title | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
title_full | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
title_fullStr | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
title_full_unstemmed | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
title_short | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
title_sort | A combined approach of rolling-circle amplification-single site restriction endonuclease digestion followed by next generation sequencing to characterize the whole genome and intra-host variants of human Torque teno virus |
topic | Torque Teno Virus Whole genome Rolling circle-amplification Restriction endonuclease digestion Next generation sequencing Intra-host variants |
url | https://hdl.handle.net/20.500.12008/42749 |