Molecular determination of Mycobacterium leprae viability by use of real-time PCR

Detalhes bibliográficos
Autor(a) principal: Martinez, Alejandra N.
Data de Publicação: 2009
Outros Autores: Lahiri, Ramanuj, Pittman, Tana L., Scollard, David, Truman, Richard, Moraes, Milton O., Williams, Diana L.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/29046
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hanseníase. Rio de Janeiro, RJ. Brasil / Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.
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spelling Martinez, Alejandra N.Lahiri, RamanujPittman, Tana L.Scollard, DavidTruman, RichardMoraes, Milton O.Williams, Diana L.2018-09-25T16:31:02Z2018-09-25T16:31:02Z2009MARTINEZ, Alejandra N. et al. Molecular Determination of Mycobacterium leprae Viability by Use of Real-Time PCR. Journal of Clinical Microbiology, v.47, n.7, p.2124-2130, July 2009.0095-1137https://www.arca.fiocruz.br/handle/icict/2904610.1128/JCM.00512-091098-660XengAmerican Society for MicrobiologyHanseníaseMycobacterium lepraeReação em Cadeia da Polimerase em Tempo Realdeterminação molecularLeprosyMycobacterium lepraeReal-Time PCRMolecular DeterminationMolecular determination of Mycobacterium leprae viability by use of real-time PCRinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hanseníase. Rio de Janeiro, RJ. Brasil / Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hanseníase. Rio de Janeiro, RJ. Brasil.Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.Mycobacterium leprae, the etiological agent of leprosy, is noncultivable on axenic media. Therefore, the viability of M. leprae for clinical or experimental applications is often unknown. To provide new tools for M. leprae viability determination, two quantitative reverse transcriptase PCR (RT-PCR) assays were developed and characterized. M. leprae sodA mRNA and 16S rRNA were used as RNA targets, and M. leprae repetitive element (RLEP) DNA was used to determine relative bacterial numbers in the same purified bacterial preparations or from crude biological specimens. Results demonstrated that both assays were good predictors of M. leprae viability during short-term experiments (48 h) involving rifampin (rifampicin) treatment in axenic medium, within rifampin-treated murine macrophages (MPhi), or within immune-activated MPhi. Moreover, these results strongly correlated those of other M. leprae viability assays, including radiorespirometry-based and Live/Dead BacLight viability assays. The 16S rRNA/RLEP assay consistently identified the presence of M. leprae in eight multibacillary leprosy patient biopsy specimens prior to multidrug therapy (MDT) and demonstrated a decline in viability during the course of MDT. In contrast, the sodA/RLEP assay was able to detect the presence of M. leprae in only 25% of pretreatment biopsy specimens. In conclusion, new tools for M. leprae viability determination were developed. The 16S rRNA/RLEP RT-PCR M. leprae viability assay should be useful both for short-term experimental purposes and for predicting M. leprae viability in biopsy specimens to monitor treatment efficacy, whereas the sodA/RLEP RT-PCR M. leprae viability assay should be limited to short-term experimental research purposes.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/29046/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALmiltonO_moraes_etal_IOC_2009.pdfmiltonO_moraes_etal_IOC_2009.pdfapplication/pdf277038https://www.arca.fiocruz.br/bitstream/icict/29046/2/miltonO_moraes_etal_IOC_2009.pdf48d699b1b8499d9bce94688f69a3f082MD52TEXTmiltonO_moraes_etal_IOC_2009.pdf.txtmiltonO_moraes_etal_IOC_2009.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Molecular determination of Mycobacterium leprae viability by use of real-time PCR
title Molecular determination of Mycobacterium leprae viability by use of real-time PCR
spellingShingle Molecular determination of Mycobacterium leprae viability by use of real-time PCR
Martinez, Alejandra N.
Hanseníase
Mycobacterium leprae
Reação em Cadeia da Polimerase em Tempo Real
determinação molecular
Leprosy
Mycobacterium leprae
Real-Time PCR
Molecular Determination
title_short Molecular determination of Mycobacterium leprae viability by use of real-time PCR
title_full Molecular determination of Mycobacterium leprae viability by use of real-time PCR
title_fullStr Molecular determination of Mycobacterium leprae viability by use of real-time PCR
title_full_unstemmed Molecular determination of Mycobacterium leprae viability by use of real-time PCR
title_sort Molecular determination of Mycobacterium leprae viability by use of real-time PCR
author Martinez, Alejandra N.
author_facet Martinez, Alejandra N.
Lahiri, Ramanuj
Pittman, Tana L.
Scollard, David
Truman, Richard
Moraes, Milton O.
Williams, Diana L.
author_role author
author2 Lahiri, Ramanuj
Pittman, Tana L.
Scollard, David
Truman, Richard
Moraes, Milton O.
Williams, Diana L.
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Martinez, Alejandra N.
Lahiri, Ramanuj
Pittman, Tana L.
Scollard, David
Truman, Richard
Moraes, Milton O.
Williams, Diana L.
dc.subject.other.pt_BR.fl_str_mv Hanseníase
Mycobacterium leprae
Reação em Cadeia da Polimerase em Tempo Real
determinação molecular
topic Hanseníase
Mycobacterium leprae
Reação em Cadeia da Polimerase em Tempo Real
determinação molecular
Leprosy
Mycobacterium leprae
Real-Time PCR
Molecular Determination
dc.subject.en.pt_BR.fl_str_mv Leprosy
Mycobacterium leprae
Real-Time PCR
Molecular Determination
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hanseníase. Rio de Janeiro, RJ. Brasil / Louisiana State University. Laboratory Research Branch at the School of Veterinary Medicine. Division of National Hansen’s Disease Programs,. HRSA, BPHC. Louisiana, USA.
publishDate 2009
dc.date.issued.fl_str_mv 2009
dc.date.accessioned.fl_str_mv 2018-09-25T16:31:02Z
dc.date.available.fl_str_mv 2018-09-25T16:31:02Z
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 MARTINEZ, Alejandra N. et al. Molecular Determination of Mycobacterium leprae Viability by Use of Real-Time PCR. Journal of Clinical Microbiology, v.47, n.7, p.2124-2130, July 2009.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/29046
dc.identifier.issn.pt_BR.fl_str_mv 0095-1137
dc.identifier.doi.none.fl_str_mv 10.1128/JCM.00512-09
dc.identifier.eissn.none.fl_str_mv 1098-660X
identifier_str_mv MARTINEZ, Alejandra N. et al. Molecular Determination of Mycobacterium leprae Viability by Use of Real-Time PCR. Journal of Clinical Microbiology, v.47, n.7, p.2124-2130, July 2009.
0095-1137
10.1128/JCM.00512-09
1098-660X
url https://www.arca.fiocruz.br/handle/icict/29046
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.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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