Multilocus Sequence Typing Scheme versus Pulsed-Field Gel Electrophoresis for Typing Mycobacterium abscessus Isolates

Author(s):Show less -

Machado, Gabriel Esquitini ^[1] ; Matsumoto, Cristianne Kayoko ^[1] ; Chimara, Erica ^[2] ; Duarte, Rafael da Silva ^[3] ; de Freitas, Denise ^[4] ; Palaci, Moises ^[5] ; Hadad, David Jamil ^[5] ; Batista Lima, Karla Valeria ^[6] ; Lopes, Maria Luiza ^[6] ; Ramos, Jesus Pais ^[7] ; Campos, Carlos Eduardo ^[7] ; Caldas, Paulo Cesar ^[7] ; Heym, Beate ^[8] ; Leao, Sylvia Cardoso ^[1]

Affiliation

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo - Brazil

^[2] Adolfo Lutz Inst, Nucleo TB & Micobacterioses, Sao Paulo - Brazil

^[3] Univ Fed Rio de Janeiro, Inst Microbiol Paulo de Goes, Rio De Janeiro - Brazil

^[4] Univ Fed Sao Paulo, Escola Paulista Med, Dept Oftalmol, Sao Paulo - Brazil

^[5] Univ Fed Espirito Santo, Nucleo Doencas Infecciosas, Vitoria, ES - Brazil

^[6] Inst Evandro Chagas, Belem, Para - Brazil

^[7] Fiocruz MS, Escola Nacl Saude Publ, Ctr Referencia Prof Helio Fraga, BR-21045900 Rio De Janeiro - Brazil

^[8] Hop Univ Paris Ile de France Ouest, Hop Ambroise Pare, AP HP, Microbiol Lab, Boulogne - France

• Document type: Journal article
• Source: Journal of Clinical Microbiology; v. 52, n. 8, p. 2881-2891, AUG 2014.
• Web of Science Citations: 6

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. (AU)

• FAPESP's process: 11/18326-4 - Mycobacteria and extrachromosomal elements: molecular characterization and biotechnological applications
• Grantee: Sylvia Luisa Pincherle Cardoso Leão
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 13/16018-6 - Sequencing and annotation of the genome from strain EPM 10906, a new mycobacterium species
• Grantee: Cristianne Kayoko Matsumoto
• Support Opportunities: Scholarships in Brazil - Post-Doctoral