title: Multilocus sequence typing scheme versus pulsed-field gel electrophoresis for typing Mycobacterium abscessus isolates.
abstract: Outbreaks of infections by rapidly growing mycobacteria following invasive procedures, such as ophthalmological, laparoscopic, arthroscopic, plastic, and cardiac surgeries, mesotherapy, and vaccination, have been detected in Brazil since 1998. Members of the Mycobacterium chelonae-Mycobacterium abscessus group have caused most of these outbreaks. As part of an epidemiological investigation, the isolates were typed by pulsed-field gel electrophoresis (PFGE). In this project, we performed a large-scale comparison of PFGE profiles with the results of a recently developed multilocus sequence typing (MLST) scheme for M. abscessus. Ninety-three isolates were analyzed, with 40 M. abscessus subsp. abscessus isolates, 47 M. abscessus subsp. bolletii isolates, and six isolates with no assigned subspecies. Forty-five isolates were obtained during five outbreaks, and 48 were sporadic isolates that were not associated with outbreaks. For MLST, seven housekeeping genes (argH, cya, glpK, gnd, murC, pta, and purH) were sequenced, and each isolate was assigned a sequence type (ST) from the combination of obtained alleles. The PFGE patterns of DraI-digested DNA were compared with the MLST results. All isolates were analyzable by both methods. Isolates from monoclonal outbreaks showed unique STs and indistinguishable or very similar PFGE patterns. Thirty-three STs and 49 unique PFGE patterns were identified among the 93 isolates. The Simpson's index of diversity values for MLST and PFGE were 0.69 and 0.93, respectively, for M. abscessus subsp. abscessus and 0.96 and 0.97, respectively, for M. abscessus subsp. bolletii. In conclusion, the MLST scheme showed 100% typeability and grouped monoclonal outbreak isolates in agreement with PFGE, but it was less discriminative than PFGE for M. abscessus.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Gabriel Esquitini
authors.initials: GE
authors.lastname: Machado
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Cristianne Kayoko
authors.initials: CK
authors.lastname: Matsumoto
authors.affiliation: Núcleo de Tuberculose e Micobacterioses, Instituto Adolfo Lutz, São Paulo, Brazil.
authors.forename: Erica
authors.initials: E
authors.lastname: Chimara
authors.affiliation: Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Rafael da Silva
authors.initials: Rda S
authors.lastname: Duarte
authors.affiliation: Departamento de Oftalmologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Denise
authors.initials: D
authors.lastname: de Freitas
authors.affiliation: Nucleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, Espírito Santo, Brazil.
authors.forename: Moises
authors.initials: M
authors.lastname: Palaci
authors.affiliation: Nucleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, Espírito Santo, Brazil.
authors.forename: David Jamil
authors.initials: DJ
authors.lastname: Hadad
authors.affiliation: Instituto Evandro Chagas, Belém, Pará, Brazil.
authors.forename: Karla Valéria Batista
authors.initials: KV
authors.lastname: Lima
authors.affiliation: Instituto Evandro Chagas, Belém, Pará, Brazil.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Lopes
authors.affiliation: Centro de Referência Professor Hélio Fraga, Escola Nacional de Saúde Pública, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Jesus Pais
authors.initials: JP
authors.lastname: Ramos
authors.affiliation: Centro de Referência Professor Hélio Fraga, Escola Nacional de Saúde Pública, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Carlos Eduardo
authors.initials: CE
authors.lastname: Campos
authors.affiliation: Centro de Referência Professor Hélio Fraga, Escola Nacional de Saúde Pública, Fiocruz, Rio de Janeiro, Brazil.
authors.forename: Paulo César
authors.initials: PC
authors.lastname: Caldas
authors.affiliation: APHP, Hôpitaux Universitaires Paris Ile-de-France Ouest, Laboratoire de Microbiologie, Hôpital Ambroise Paré, Boulogne-Billancourt, France.
authors.forename: Beate
authors.initials: B
authors.lastname: Heym
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil sylvia.leao@unifesp.br.
authors.forename: Sylvia Cardoso
authors.initials: SC
authors.lastname: Leão
mesh_terms: D001938:Brazil / Q000453:epidemiology; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D006801:Humans; D017720:Molecular Epidemiology / Q000379:methods; D058885:Multilocus Sequence Typing / Q000379:methods*; D009165:Mycobacterium Infections, Nontuberculous / Q000453:epidemiology; D009170:Nontuberculous Mycobacteria / Q000145:classification* / Q000235:genetics*