Rational design of diagnostic and vaccination strategies for tuberculosis
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da FURG (RI FURG) |
Texto Completo: | http://repositorio.furg.br/handle/1/7402 http://dx.doi.org/10.1590/S1413-86702012000100012 |
Resumo: | The development of diagnostic tests which can readily differentiate between vaccinated and tuberculosis-infected individuals is crucial for the wider utilization of bacillus Calmette-Guérin (BCG) as vaccine in humans and animals. BCG_0092 is an antigen that elicits specific delayed type hypersensitivity reactions similar in size and morphological aspects to that elicited by purified protein derivative, in both animals and humans infected with the tubercle bacilli. We carried out bioinformatics analyses of the BCG_0092 and designed a diagnostic test by using the predicted MHC class I epitopes. In addition, we performed a knockout of this gene by homologous recombination in the BCG vaccine strain to allow differentiation of vaccinated from infected individuals. For that, the flanking sequences of the target gene (BCG_0092)were cloned into a suicide vector. Spontaneous double crossovers, which result in wild type revertants or knockouts were selected using SacB. BCG_0092 is present only in members of the Mycobacterium tuberculosis complex. Eight predicted MHC class I epitopes with potential for immunological diagnosis were defined, allowing the design of a specific diagnostic test. The strategy used to delete the (BCG_0092) gene from BCG was successful. The knockout genotype was confirmed by PCR and by Southern blot. The mutant BCG strain has the potential of inducing protection against tuberculosis without interfering with the diagnostic test based on the use of selected epitopes from BCG_0092. |
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Borsuk, SibeleSeixas, Fabiana KömmlingSoares, Daniela Fernandes RamosMendum, TomMcFadden, JohnjoeDellagostin, Odir Antonio2017-07-19T18:22:51Z2017-07-19T18:22:51Z2012BORSUK, Sibele et al. Rational design of diagnostic and vaccination strategies for tuberculosis. The Brazilian Journal of Infectious Diseases, v. 16, n. 1, p. 68-73, 2012. Disponível em:< http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702012000100012&lng=en&nrm=iso&tlng=en>. Acesso em: 09 abr. 2017.1413-8670http://repositorio.furg.br/handle/1/7402http://dx.doi.org/10.1590/S1413-86702012000100012The development of diagnostic tests which can readily differentiate between vaccinated and tuberculosis-infected individuals is crucial for the wider utilization of bacillus Calmette-Guérin (BCG) as vaccine in humans and animals. BCG_0092 is an antigen that elicits specific delayed type hypersensitivity reactions similar in size and morphological aspects to that elicited by purified protein derivative, in both animals and humans infected with the tubercle bacilli. We carried out bioinformatics analyses of the BCG_0092 and designed a diagnostic test by using the predicted MHC class I epitopes. In addition, we performed a knockout of this gene by homologous recombination in the BCG vaccine strain to allow differentiation of vaccinated from infected individuals. For that, the flanking sequences of the target gene (BCG_0092)were cloned into a suicide vector. Spontaneous double crossovers, which result in wild type revertants or knockouts were selected using SacB. BCG_0092 is present only in members of the Mycobacterium tuberculosis complex. Eight predicted MHC class I epitopes with potential for immunological diagnosis were defined, allowing the design of a specific diagnostic test. The strategy used to delete the (BCG_0092) gene from BCG was successful. The knockout genotype was confirmed by PCR and by Southern blot. The mutant BCG strain has the potential of inducing protection against tuberculosis without interfering with the diagnostic test based on the use of selected epitopes from BCG_0092.engGene knockout techniquesClinical diagnosisBCG vaccineRational design of diagnostic and vaccination strategies for tuberculosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FURG (RI FURG)instname:Universidade Federal do Rio Grande (FURG)instacron:FURGORIGINALv16n1a12.pdfv16n1a12.pdfapplication/pdf877855https://repositorio.furg.br/bitstream/1/7402/1/v16n1a12.pdffffb03892a04e98cd7983cb816c63e96MD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.furg.br/bitstream/1/7402/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52open access1/74022022-10-21 16:42:39.29open accessoai:repositorio.furg.br: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Repositório InstitucionalPUBhttps://repositorio.furg.br/oai/request || http://200.19.254.174/oai/requestopendoar:2022-10-21T19:42:39Repositório Institucional da FURG (RI FURG) - Universidade Federal do Rio Grande (FURG)false |
dc.title.pt_BR.fl_str_mv |
Rational design of diagnostic and vaccination strategies for tuberculosis |
title |
Rational design of diagnostic and vaccination strategies for tuberculosis |
spellingShingle |
Rational design of diagnostic and vaccination strategies for tuberculosis Borsuk, Sibele Gene knockout techniques Clinical diagnosis BCG vaccine |
title_short |
Rational design of diagnostic and vaccination strategies for tuberculosis |
title_full |
Rational design of diagnostic and vaccination strategies for tuberculosis |
title_fullStr |
Rational design of diagnostic and vaccination strategies for tuberculosis |
title_full_unstemmed |
Rational design of diagnostic and vaccination strategies for tuberculosis |
title_sort |
Rational design of diagnostic and vaccination strategies for tuberculosis |
author |
Borsuk, Sibele |
author_facet |
Borsuk, Sibele Seixas, Fabiana Kömmling Soares, Daniela Fernandes Ramos Mendum, Tom McFadden, Johnjoe Dellagostin, Odir Antonio |
author_role |
author |
author2 |
Seixas, Fabiana Kömmling Soares, Daniela Fernandes Ramos Mendum, Tom McFadden, Johnjoe Dellagostin, Odir Antonio |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Borsuk, Sibele Seixas, Fabiana Kömmling Soares, Daniela Fernandes Ramos Mendum, Tom McFadden, Johnjoe Dellagostin, Odir Antonio |
dc.subject.por.fl_str_mv |
Gene knockout techniques Clinical diagnosis BCG vaccine |
topic |
Gene knockout techniques Clinical diagnosis BCG vaccine |
description |
The development of diagnostic tests which can readily differentiate between vaccinated and tuberculosis-infected individuals is crucial for the wider utilization of bacillus Calmette-Guérin (BCG) as vaccine in humans and animals. BCG_0092 is an antigen that elicits specific delayed type hypersensitivity reactions similar in size and morphological aspects to that elicited by purified protein derivative, in both animals and humans infected with the tubercle bacilli. We carried out bioinformatics analyses of the BCG_0092 and designed a diagnostic test by using the predicted MHC class I epitopes. In addition, we performed a knockout of this gene by homologous recombination in the BCG vaccine strain to allow differentiation of vaccinated from infected individuals. For that, the flanking sequences of the target gene (BCG_0092)were cloned into a suicide vector. Spontaneous double crossovers, which result in wild type revertants or knockouts were selected using SacB. BCG_0092 is present only in members of the Mycobacterium tuberculosis complex. Eight predicted MHC class I epitopes with potential for immunological diagnosis were defined, allowing the design of a specific diagnostic test. The strategy used to delete the (BCG_0092) gene from BCG was successful. The knockout genotype was confirmed by PCR and by Southern blot. The mutant BCG strain has the potential of inducing protection against tuberculosis without interfering with the diagnostic test based on the use of selected epitopes from BCG_0092. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012 |
dc.date.accessioned.fl_str_mv |
2017-07-19T18:22:51Z |
dc.date.available.fl_str_mv |
2017-07-19T18:22:51Z |
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.citation.fl_str_mv |
BORSUK, Sibele et al. Rational design of diagnostic and vaccination strategies for tuberculosis. The Brazilian Journal of Infectious Diseases, v. 16, n. 1, p. 68-73, 2012. Disponível em:< http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702012000100012&lng=en&nrm=iso&tlng=en>. Acesso em: 09 abr. 2017. |
dc.identifier.uri.fl_str_mv |
http://repositorio.furg.br/handle/1/7402 |
dc.identifier.issn.none.fl_str_mv |
1413-8670 |
dc.identifier.doi.pt_BR.fl_str_mv |
http://dx.doi.org/10.1590/S1413-86702012000100012 |
identifier_str_mv |
BORSUK, Sibele et al. Rational design of diagnostic and vaccination strategies for tuberculosis. The Brazilian Journal of Infectious Diseases, v. 16, n. 1, p. 68-73, 2012. Disponível em:< http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702012000100012&lng=en&nrm=iso&tlng=en>. Acesso em: 09 abr. 2017. 1413-8670 |
url |
http://repositorio.furg.br/handle/1/7402 http://dx.doi.org/10.1590/S1413-86702012000100012 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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reponame:Repositório Institucional da FURG (RI FURG) instname:Universidade Federal do Rio Grande (FURG) instacron:FURG |
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