title: Diversity of pulsed-field gel electrophoresis patterns of Mycobacterium abscessus type 2 clinical isolates.
abstract: An epidemic of infections by rapidly growing mycobacteria related to surgical procedures between 2004 and 2008 in Brazil was caused by a unique strain showing the Mycobacterium abscessus type 2 pattern when it was analyzed by the molecular method of PCR-restriction enzyme analysis of the hsp65 gene (PRA-hsp65). In order to investigate the diversity of M. abscessus type 2 clinical isolates and to assess whether this epidemic strain was present in specimens from nonsurgical patients, we studied 52 isolates from 38 patients showing this characteristic PRA-hsp65 pattern obtained between 2005 and 2009. All isolates were identified by sequencing of region V of the rpoB gene and typed by pulsed-field gel electrophoresis (PFGE) using two restriction enzymes, DraI and AseI. Seven isolates obtained from sputum, bronchoalveolar lavage fluid, and urine in three different Brazilian states showed rpoB sequences 100% similar to the rpoB sequence of epidemic strain INCQS 594 and PFGE patterns highly related to the patterns of isolates, evidencing the presence of the epidemic strain in isolates from patients not associated with the surgical epidemic. The remaining isolates showed diverse rpoB sequences, with the highest similarities being to the corresponding sequences of M. massiliense(T) CIP 108297 (21 isolates), M. bolletii(T) CIP 108541 (19 isolates), or M. abscessus(T) ATCC 19977 (5 isolates). Two additional clusters could be detected by PFGE. PFGE showed 100% typeability and reproducibility and discriminatory powers, calculated by Simpson's index of diversity, of 0.978 (DraI) and 0.986 (AseI), confirming its suitability for the discrimination of M. abscessus type 2 isolates.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu, São Paulo, SP, Brazil.
authors.forename: Cristianne Kayoko
authors.initials: CK
authors.lastname: Matsumoto
authors.forename: Erica
authors.initials: E
authors.lastname: Chimara
authors.forename: Sidney
authors.initials: S
authors.lastname: Bombarda
authors.forename: Rafael Silva
authors.initials: RS
authors.lastname: Duarte
authors.forename: Sylvia Cardoso
authors.initials: SC
authors.lastname: Leão
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics; D001938:Brazil; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D018834:Chaperonin 60 / Q000235:genetics; D016000:Cluster Analysis; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D058889:Molecular Typing; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013183:Sputum / Q000382:microbiology; D014556:Urine / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; C074119:heat-shock protein 65, Mycobacterium; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit