Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease

Detalhes bibliográficos
Autor(a) principal: Sturm, Nancy R.
Data de Publicação: 1989
Outros Autores: Degrave, Wim, Morel, Carlos Medicis, Simpson, Larry
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/14600
Resumo: University of California. Department of Biology and Molecular Biology Institute. Los Angeles, CA, USA.
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spelling Sturm, Nancy R.Degrave, WimMorel, Carlos MedicisSimpson, Larry2016-06-29T13:25:05Z2016-07-07T11:40:25Z2016-06-29T13:25:05Z2016-07-07T11:40:25Z1989STURM, Nancy R. et al. Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease. Molecular and Biochemical Parasitology, v. 33, p. 205-214, 1989.0166-6851https://www.arca.fiocruz.br/handle/icict/1460010.1016/0166-6851(89)90082-01872-9428engElsevierSensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' diseaseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversity of California. Department of Biology and Molecular Biology Institute. Los Angeles, CA, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Rio de Janeiro, RJ, BrasilUniversity of California. Department of Biology and Molecular Biology Institute. Los Angeles, CA, USA.Amplification of DNA sequences from the kinetoplast minicircle DNA was employed as a method for the detection and classification of small numbers of Trypanosoma cruzi cells. Two overlapping fragments from the conserved 120 bp minirepeat regions of the minicircle DNA and one fragment covering the adjacent variable regions were amplified. The minimal amount of minicircle DNA required to detect a product by hybridization with an oligonucleotide probe was 0.015 fg. which represents approximately 10 molecules or 0.1% of the minicircle DNA component of a single cell. The amplification worked equally well with kDNA from several strains of T. cruzi and did not occur with kDNA from several other kinetoplastids. kDNA recovered from less than 10 trypanosomes in whole blood could be used as a template for amplification; the presence of a several billion fold excess of human DNA had no effect on the amplification process. Schizodeme analysis by hybridization with specific oligonucleotides or by direct restriction enzyme digestion could be performed on the amplified fragments representing the minicircle conserved region or variable regions. This method should prove useful as a rapid, specific and sensitive assay for Chagas' disease in chronic patients as well as for epidemiological studies of infected animals and insects.Trypanosoma cruziChagas DiseasePolymerase chain reactionMinirepeatMinicircle DNASchizodemeDNA de CinetoplastoTrypanosoma cruziDoença de ChagasReação em Cadeia da Polimeraseinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZTEXTcarlos11_morel_etal_IOC_1989.pdf.txtcarlos11_morel_etal_IOC_1989.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
title Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
spellingShingle Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
Sturm, Nancy R.
Trypanosoma cruzi
Chagas Disease
Polymerase chain reaction
Minirepeat
Minicircle DNA
Schizodeme
DNA de Cinetoplasto
Trypanosoma cruzi
Doença de Chagas
Reação em Cadeia da Polimerase
title_short Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
title_full Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
title_fullStr Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
title_full_unstemmed Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
title_sort Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease
author Sturm, Nancy R.
author_facet Sturm, Nancy R.
Degrave, Wim
Morel, Carlos Medicis
Simpson, Larry
author_role author
author2 Degrave, Wim
Morel, Carlos Medicis
Simpson, Larry
author2_role author
author
author
dc.contributor.author.fl_str_mv Sturm, Nancy R.
Degrave, Wim
Morel, Carlos Medicis
Simpson, Larry
dc.subject.en.pt_BR.fl_str_mv Trypanosoma cruzi
Chagas Disease
Polymerase chain reaction
Minirepeat
Minicircle DNA
Schizodeme
topic Trypanosoma cruzi
Chagas Disease
Polymerase chain reaction
Minirepeat
Minicircle DNA
Schizodeme
DNA de Cinetoplasto
Trypanosoma cruzi
Doença de Chagas
Reação em Cadeia da Polimerase
dc.subject.decs.pt_BR.fl_str_mv DNA de Cinetoplasto
Trypanosoma cruzi
Doença de Chagas
Reação em Cadeia da Polimerase
description University of California. Department of Biology and Molecular Biology Institute. Los Angeles, CA, USA.
publishDate 1989
dc.date.issued.fl_str_mv 1989
dc.date.accessioned.fl_str_mv 2016-06-29T13:25:05Z
2016-07-07T11:40:25Z
dc.date.available.fl_str_mv 2016-06-29T13:25:05Z
2016-07-07T11:40:25Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.citation.fl_str_mv STURM, Nancy R. et al. Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease. Molecular and Biochemical Parasitology, v. 33, p. 205-214, 1989.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/14600
dc.identifier.issn.pt_BR.fl_str_mv 0166-6851
dc.identifier.doi.pt_BR.fl_str_mv 10.1016/0166-6851(89)90082-0
dc.identifier.eissn.pt_BR.fl_str_mv 1872-9428
identifier_str_mv STURM, Nancy R. et al. Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease. Molecular and Biochemical Parasitology, v. 33, p. 205-214, 1989.
0166-6851
10.1016/0166-6851(89)90082-0
1872-9428
url https://www.arca.fiocruz.br/handle/icict/14600
dc.language.iso.fl_str_mv eng
language eng
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dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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instacron_str FIOCRUZ
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collection Repositório Institucional da FIOCRUZ (ARCA)
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