Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.

Detalhes bibliográficos
Autor(a) principal: Pimenta, Juliana Ramos
Data de Publicação: 2008
Outros Autores: Freitas, Jorge Marcelo de, Duffy, Tomás, Bartholomeu, Daniella Castanheira, Oliveira, Riva de Paula, Chiari, Egler, Moreira, Maria da Consolação Vieira, Brasileiro Filho, Geraldo, Schijman, Alejandro Gabriel, Franco, Glória Regina, Machado, Carlos Renato, Pena, Sérgio Danilo Junho, Macedo, Andréa Mara
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFOP
Texto Completo: http://www.repositorio.ufop.br/handle/123456789/4798
https://doi.org/10.1016/j.ijpara.2007.10.017
Resumo: The investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci – six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.
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spelling Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.Trypanosoma cruziChagas diseaseGenome projectPolymorphic microsatellitesFull nestedThe investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci – six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.2015-03-31T17:21:29Z2015-03-31T17:21:29Z2008info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfPIMENTA, J. R. et al. Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites. International Journal for Parasitology, v. 38, p. 839-850, 2008. Disponível em: <http://www.sciencedirect.com/science/article/pii/S0020751907003918>. Acesso em: 15 out. 2014.0020-7519http://www.repositorio.ufop.br/handle/123456789/4798https://doi.org/10.1016/j.ijpara.2007.10.017O periódico International Journal for Parasitology concede permissão para depósito deste artigo no Repositório Institucional da UFOP. Número da licença: 3493711276878.info:eu-repo/semantics/openAccessPimenta, Juliana RamosFreitas, Jorge Marcelo deDuffy, TomásBartholomeu, Daniella CastanheiraOliveira, Riva de PaulaChiari, EglerMoreira, Maria da Consolação VieiraBrasileiro Filho, GeraldoSchijman, Alejandro GabrielFranco, Glória ReginaMachado, Carlos RenatoPena, Sérgio Danilo JunhoMacedo, Andréa Maraengreponame:Repositório Institucional da UFOPinstname:Universidade Federal de Ouro Preto (UFOP)instacron:UFOP2019-07-01T12:57:34Zoai:repositorio.ufop.br:123456789/4798Repositório InstitucionalPUBhttp://www.repositorio.ufop.br/oai/requestrepositorio@ufop.edu.bropendoar:32332019-07-01T12:57:34Repositório Institucional da UFOP - Universidade Federal de Ouro Preto (UFOP)false
dc.title.none.fl_str_mv Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
title Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
spellingShingle Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
Pimenta, Juliana Ramos
Trypanosoma cruzi
Chagas disease
Genome project
Polymorphic microsatellites
Full nested
title_short Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
title_full Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
title_fullStr Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
title_full_unstemmed Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
title_sort Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
author Pimenta, Juliana Ramos
author_facet Pimenta, Juliana Ramos
Freitas, Jorge Marcelo de
Duffy, Tomás
Bartholomeu, Daniella Castanheira
Oliveira, Riva de Paula
Chiari, Egler
Moreira, Maria da Consolação Vieira
Brasileiro Filho, Geraldo
Schijman, Alejandro Gabriel
Franco, Glória Regina
Machado, Carlos Renato
Pena, Sérgio Danilo Junho
Macedo, Andréa Mara
author_role author
author2 Freitas, Jorge Marcelo de
Duffy, Tomás
Bartholomeu, Daniella Castanheira
Oliveira, Riva de Paula
Chiari, Egler
Moreira, Maria da Consolação Vieira
Brasileiro Filho, Geraldo
Schijman, Alejandro Gabriel
Franco, Glória Regina
Machado, Carlos Renato
Pena, Sérgio Danilo Junho
Macedo, Andréa Mara
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Pimenta, Juliana Ramos
Freitas, Jorge Marcelo de
Duffy, Tomás
Bartholomeu, Daniella Castanheira
Oliveira, Riva de Paula
Chiari, Egler
Moreira, Maria da Consolação Vieira
Brasileiro Filho, Geraldo
Schijman, Alejandro Gabriel
Franco, Glória Regina
Machado, Carlos Renato
Pena, Sérgio Danilo Junho
Macedo, Andréa Mara
dc.subject.por.fl_str_mv Trypanosoma cruzi
Chagas disease
Genome project
Polymorphic microsatellites
Full nested
topic Trypanosoma cruzi
Chagas disease
Genome project
Polymorphic microsatellites
Full nested
description The investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci – six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.
publishDate 2008
dc.date.none.fl_str_mv 2008
2015-03-31T17:21:29Z
2015-03-31T17:21:29Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv PIMENTA, J. R. et al. Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites. International Journal for Parasitology, v. 38, p. 839-850, 2008. Disponível em: <http://www.sciencedirect.com/science/article/pii/S0020751907003918>. Acesso em: 15 out. 2014.
0020-7519
http://www.repositorio.ufop.br/handle/123456789/4798
https://doi.org/10.1016/j.ijpara.2007.10.017
identifier_str_mv PIMENTA, J. R. et al. Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites. International Journal for Parasitology, v. 38, p. 839-850, 2008. Disponível em: <http://www.sciencedirect.com/science/article/pii/S0020751907003918>. Acesso em: 15 out. 2014.
0020-7519
url http://www.repositorio.ufop.br/handle/123456789/4798
https://doi.org/10.1016/j.ijpara.2007.10.017
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFOP
instname:Universidade Federal de Ouro Preto (UFOP)
instacron:UFOP
instname_str Universidade Federal de Ouro Preto (UFOP)
instacron_str UFOP
institution UFOP
reponame_str Repositório Institucional da UFOP
collection Repositório Institucional da UFOP
repository.name.fl_str_mv Repositório Institucional da UFOP - Universidade Federal de Ouro Preto (UFOP)
repository.mail.fl_str_mv repositorio@ufop.edu.br
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