Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

Detalhes bibliográficos
Autor(a) principal: Michelon, Candice Tosi
Data de Publicação: 2011
Outros Autores: Rosso, Franciele, Schmid, Karen Barros, Sperhacke, Rosa Dea, Oliveira, Martha Maria, Kritski, Afrânio Lineu, Rezende Junior, Leonides, Costa, Elis Regina Dalla, Ribeiro, Andrezza Wolowski, Verza, Mirela, Cafrune, Patricia Izquierdo, Silva, Marcia Susana Nunes, Kühleis, Daniele Chaves, Zaha, Arnaldo, Rossetti, Maria Lucia Rosa
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFRGS
Texto Completo: http://hdl.handle.net/10183/30533
Resumo: Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.
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spelling Michelon, Candice TosiRosso, FrancieleSchmid, Karen BarrosSperhacke, Rosa DeaOliveira, Martha MariaKritski, Afrânio LineuRezende Junior, LeonidesCosta, Elis Regina DallaRibeiro, Andrezza WolowskiVerza, MirelaCafrune, Patricia IzquierdoSilva, Marcia Susana NunesKühleis, Daniele ChavesZaha, ArnaldoRossetti, Maria Lucia Rosa2011-08-04T06:01:08Z20110074-0276http://hdl.handle.net/10183/30533000776733Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.application/pdfengMemorias do Instituto Oswaldo Cruz : an International Journal of Biological and Biomedical Research. Rio de Janeiro. Vol. 106, n. 2 (mar. 2011), p. 194-199Mycobacterium tuberculosisTuberculoseHibridizaçãoTuberculosisReverseHybridization assayIS6110Molecular diagnosisOligonucleotide probeColorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detectioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/otherinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSORIGINAL000776733.pdf000776733.pdfTexto completo (inglês)application/pdf332837http://www.lume.ufrgs.br/bitstream/10183/30533/1/000776733.pdfabb6b349c9523f537f0046099be60994MD51TEXT000776733.pdf.txt000776733.pdf.txtExtracted Texttext/plain34650http://www.lume.ufrgs.br/bitstream/10183/30533/2/000776733.pdf.txt85fbae56cb9123d31e92789429a5cec2MD52THUMBNAIL000776733.pdf.jpg000776733.pdf.jpgGenerated Thumbnailimage/jpeg2206http://www.lume.ufrgs.br/bitstream/10183/30533/3/000776733.pdf.jpg7b8bbae9e0e22383177484961be8a766MD5310183/305332018-10-15 09:24:59.167oai:www.lume.ufrgs.br:10183/30533Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2018-10-15T12:24:59Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false
dc.title.pt_BR.fl_str_mv Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
title Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
spellingShingle Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
Michelon, Candice Tosi
Mycobacterium tuberculosis
Tuberculose
Hibridização
Tuberculosis
Reverse
Hybridization assay
IS6110
Molecular diagnosis
Oligonucleotide probe
title_short Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
title_full Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
title_fullStr Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
title_full_unstemmed Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
title_sort Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
author Michelon, Candice Tosi
author_facet Michelon, Candice Tosi
Rosso, Franciele
Schmid, Karen Barros
Sperhacke, Rosa Dea
Oliveira, Martha Maria
Kritski, Afrânio Lineu
Rezende Junior, Leonides
Costa, Elis Regina Dalla
Ribeiro, Andrezza Wolowski
Verza, Mirela
Cafrune, Patricia Izquierdo
Silva, Marcia Susana Nunes
Kühleis, Daniele Chaves
Zaha, Arnaldo
Rossetti, Maria Lucia Rosa
author_role author
author2 Rosso, Franciele
Schmid, Karen Barros
Sperhacke, Rosa Dea
Oliveira, Martha Maria
Kritski, Afrânio Lineu
Rezende Junior, Leonides
Costa, Elis Regina Dalla
Ribeiro, Andrezza Wolowski
Verza, Mirela
Cafrune, Patricia Izquierdo
Silva, Marcia Susana Nunes
Kühleis, Daniele Chaves
Zaha, Arnaldo
Rossetti, Maria Lucia Rosa
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Michelon, Candice Tosi
Rosso, Franciele
Schmid, Karen Barros
Sperhacke, Rosa Dea
Oliveira, Martha Maria
Kritski, Afrânio Lineu
Rezende Junior, Leonides
Costa, Elis Regina Dalla
Ribeiro, Andrezza Wolowski
Verza, Mirela
Cafrune, Patricia Izquierdo
Silva, Marcia Susana Nunes
Kühleis, Daniele Chaves
Zaha, Arnaldo
Rossetti, Maria Lucia Rosa
dc.subject.por.fl_str_mv Mycobacterium tuberculosis
Tuberculose
Hibridização
topic Mycobacterium tuberculosis
Tuberculose
Hibridização
Tuberculosis
Reverse
Hybridization assay
IS6110
Molecular diagnosis
Oligonucleotide probe
dc.subject.eng.fl_str_mv Tuberculosis
Reverse
Hybridization assay
IS6110
Molecular diagnosis
Oligonucleotide probe
description Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.
publishDate 2011
dc.date.accessioned.fl_str_mv 2011-08-04T06:01:08Z
dc.date.issued.fl_str_mv 2011
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dc.language.iso.fl_str_mv eng
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dc.relation.ispartof.pt_BR.fl_str_mv Memorias do Instituto Oswaldo Cruz : an International Journal of Biological and Biomedical Research. Rio de Janeiro. Vol. 106, n. 2 (mar. 2011), p. 194-199
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