Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

Abstract

Submitted by Nuzia Santos ([email protected]) on 2015-02-27T12:50:33Z No. of bitstreams: 1 2014_112.pdf: 385803 bytes, checksum: 73ae8e5baf37fce92c3d713b7f570aed (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-27T12:50:39Z (GMT) No. of bitstreams: 1 2014_112.pdf: 385803 bytes, checksum: 73ae8e5baf37fce92c3d713b7f570aed (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-27T12:56:22Z (GMT) No. of bitstreams: 1 2014_112.pdf: 385803 bytes, checksum: 73ae8e5baf37fce92c3d713b7f570aed (MD5)Made available in DSpace on 2015-02-27T12:56:22Z (GMT). No. of bitstreams: 1 2014_112.pdf: 385803 bytes, checksum: 73ae8e5baf37fce92c3d713b7f570aed (MD5) Previous issue date: 2014CNPq, LMW and MLBUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Biologia Molecular e Micobactérias. Florianópolis, SC, BrasilUniversidade Federal de Santa Catarina. Laboratório de Protozoologia. Florianópolis, SC, BrasilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, BrasilLaboratório Central do Estado de Santa Catarina. Florianópolis, SC, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Laboratório de Vírus. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Laboratório de Vírus. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Laboratório de Vírus. Belo Horizonte, MG, BrasilThe identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteri

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