title: Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria.
abstract: The 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 mycobacteria.
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Letícia Muraro
authors.initials: LM
authors.lastname: Wildner
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Bazzo
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Susie Coutinho
authors.initials: SC
authors.lastname: Liedke
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Christiane Lourenço
authors.initials: CL
authors.lastname: Nogueira
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Segat
authors.affiliation: Laboratório de Biologia Molecular e Micobactérias, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Simone Gonçalves
authors.initials: SG
authors.lastname: Senna
authors.affiliation: Laboratório de Protozoologia, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Aline Daiane
authors.initials: AD
authors.lastname: Schlindwein
authors.affiliation: Laboratório de Imunologia Celular e Molecular, Centro de Pesquisas René Rachou, Fiocruz, Belo Horizonte, MG, Brasil.
authors.forename: Jaquelline Germano de
authors.initials: JG
authors.lastname: Oliveira
authors.affiliation: Laboratório Central, Estado de Santa Catarina, Florianópolis, SC, Brasil.
authors.forename: Darcita B
authors.initials: DB
authors.lastname: Rovaris
authors.affiliation: Laboratório de Vírus, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil.
authors.forename: Claudio A
authors.initials: CA
authors.lastname: Bonjardim
authors.affiliation: Laboratório de Vírus, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil.
authors.forename: Erna G
authors.initials: EG
authors.lastname: Kroon
authors.affiliation: Laboratório de Vírus, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil.
authors.forename: Paulo C P
authors.initials: PC
authors.lastname: Ferreira
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D009164:Mycobacterium Infections / Q000382:microbiology; D009165:Mycobacterium Infections, Nontuberculous / Q000382:microbiology; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000302:isolation & purification*; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S