Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil.
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 EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/913965 https://doi.org/10.1590/S0074-02762011000100002 |
Resumo: | A cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC. |
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Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil.OxyR pseudogeneSputum smearMolecular sequence dataMycobacterium BovisMycobacterium tuberculosispolymerase chain reactionA cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC.MARCIO ROBERTO SILVA, CNPGL; MARK DREW CROSLAND GUIMARÃES, UFMG; VANIA MARIA DE OLIVEIRA VEIGA, CNPGL; ALINE DOS SANTOS MOREIRA, Laboratório de Genômica Funcional e Bioinformática; RONALDO RODRIGUES DA COSTA, UFJF; FHEMIG; KELLY CRISTINA FERREIRA ABI-ZAID, FHEMIG; ADALGIZA DA SILVA ROCHA, FIOCRUZ; PHILIP NOEL SUFFYS, FIOCRUZ.SILVA, M. R.GUIMARÃES, M. D. C.VEIGA, V. M. de O.MOREIRA, A. dos S.COSTA, R. R. daABI-ZAID, K. C. F.ROCHA, A. da S.SUFFYS, P. N.2024-02-06T16:32:57Z2024-02-06T16:32:57Z2012-01-312011info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleMemorias do Instituto Oswaldo Cruz, v. 106, n. 1, p. 9-15, 2011.http://www.alice.cnptia.embrapa.br/alice/handle/doc/913965https://doi.org/10.1590/S0074-02762011000100002enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2024-02-06T16:32:57Zoai:www.alice.cnptia.embrapa.br:doc/913965Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542024-02-06T16:32:57falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542024-02-06T16:32:57Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
title |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
spellingShingle |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. SILVA, M. R. OxyR pseudogene Sputum smear Molecular sequence data Mycobacterium Bovis Mycobacterium tuberculosis polymerase chain reaction |
title_short |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
title_full |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
title_fullStr |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
title_full_unstemmed |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
title_sort |
Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil. |
author |
SILVA, M. R. |
author_facet |
SILVA, M. R. GUIMARÃES, M. D. C. VEIGA, V. M. de O. MOREIRA, A. dos S. COSTA, R. R. da ABI-ZAID, K. C. F. ROCHA, A. da S. SUFFYS, P. N. |
author_role |
author |
author2 |
GUIMARÃES, M. D. C. VEIGA, V. M. de O. MOREIRA, A. dos S. COSTA, R. R. da ABI-ZAID, K. C. F. ROCHA, A. da S. SUFFYS, P. N. |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
MARCIO ROBERTO SILVA, CNPGL; MARK DREW CROSLAND GUIMARÃES, UFMG; VANIA MARIA DE OLIVEIRA VEIGA, CNPGL; ALINE DOS SANTOS MOREIRA, Laboratório de Genômica Funcional e Bioinformática; RONALDO RODRIGUES DA COSTA, UFJF; FHEMIG; KELLY CRISTINA FERREIRA ABI-ZAID, FHEMIG; ADALGIZA DA SILVA ROCHA, FIOCRUZ; PHILIP NOEL SUFFYS, FIOCRUZ. |
dc.contributor.author.fl_str_mv |
SILVA, M. R. GUIMARÃES, M. D. C. VEIGA, V. M. de O. MOREIRA, A. dos S. COSTA, R. R. da ABI-ZAID, K. C. F. ROCHA, A. da S. SUFFYS, P. N. |
dc.subject.por.fl_str_mv |
OxyR pseudogene Sputum smear Molecular sequence data Mycobacterium Bovis Mycobacterium tuberculosis polymerase chain reaction |
topic |
OxyR pseudogene Sputum smear Molecular sequence data Mycobacterium Bovis Mycobacterium tuberculosis polymerase chain reaction |
description |
A cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011 2012-01-31 2024-02-06T16:32:57Z 2024-02-06T16:32:57Z |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
Memorias do Instituto Oswaldo Cruz, v. 106, n. 1, p. 9-15, 2011. http://www.alice.cnptia.embrapa.br/alice/handle/doc/913965 https://doi.org/10.1590/S0074-02762011000100002 |
identifier_str_mv |
Memorias do Instituto Oswaldo Cruz, v. 106, n. 1, p. 9-15, 2011. |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/913965 https://doi.org/10.1590/S0074-02762011000100002 |
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.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
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Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
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EMBRAPA |
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EMBRAPA |
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Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
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Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
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