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

dc.creatorMartinez, Alejandra N.
dc.creatorLahiri, Ramanuj
dc.creatorPittman, Tana L.
dc.creatorScollard, David
dc.creatorTruman, Richard
dc.creatorMoraes, Milton O.
dc.creatorWilliams, Diana L.
dc.date2018-09-25T16:31:02Z
dc.date2018-09-25T16:31:02Z
dc.date2009
dc.date.accessioned2023-09-26T20:47:30Z
dc.date.available2023-09-26T20:47:30Z
dc.identifierMARTINEZ, 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.identifier0095-1137
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/29046
dc.identifier10.1128/JCM.00512-09
dc.identifier1098-660X
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8863963
dc.descriptionMycobacterium 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherAmerican Society for Microbiology
dc.rightsopen access
dc.subjectHanseníase
dc.subjectMycobacterium leprae
dc.subjectReação em Cadeia da Polimerase em Tempo Real
dc.subjectdeterminação molecular
dc.subjectLeprosy
dc.subjectMycobacterium leprae
dc.subjectReal-Time PCR
dc.subjectMolecular Determination
dc.titleMolecular determination of Mycobacterium leprae viability by use of real-time PCR
dc.typeArticle


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