Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , , , , , , , , , , , |
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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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 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/other |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10183/30533 |
dc.identifier.issn.pt_BR.fl_str_mv |
0074-0276 |
dc.identifier.nrb.pt_BR.fl_str_mv |
000776733 |
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0074-0276 000776733 |
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http://hdl.handle.net/10183/30533 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
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