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 - Marandino, Ana - Panzera Crespo, Yanina - Bentancour, Gabriela - Mirazo, Santiago - Arbiza, Juan - Ramos, Natalia

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.


Detalles Bibliográficos
2023
Torque Teno Virus
Whole genome
Rolling circle-amplification
Restriction endonuclease digestion
Next generation sequencing
Intra-host variants
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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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.
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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
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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
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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