Molecular identification and typing of Mycobacterium massiliense isolated from postsurgical infections in Brazil

Monego, Fernanda; Duarte, Rafael Silva; Nakatani, Sueli M.; Araújo, Wildo Navegantes de; Riediger, Irina Nastassja; Brockelt, Sonia Regina; Souza, Verena Maria Mendes de; Cataldo, Jamyra Iglesias; Dias, Rubens Clayton da Silva; Biondo, Alexander Welker

doi:10.1016/s1413-8670(11)70224-0

Cited by 16 publications

(19 citation statements)

References 25 publications

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“…Outbreak 2 was a nationwide epidemic of surgical site infections related to laparoscopic and arthroscopic surgeries, which generated Ͼ2,000 notifications from several states to the Brazilian federal authorities between 2004 and 2008 (10,(31)(32)(33). The study of 157 isolates from the seven states that reported the most cases showed that the infections were caused by a single strain of M. abscessus subsp.…”

Section: Mycobacterial Isolatesmentioning

confidence: 99%

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

et al. 2014

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hOutbreaks 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.

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Section: Mycobacterial Isolatesmentioning

confidence: 99%

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

et al. 2014

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“…The M. abscessus complex comprises ubiquitous environmental micro-organisms that are frequently associated with hospital-acquired outbreaks and pseudo-outbreaks (Brown-Elliott & Wallace, 2002;Duarte et al, 2009;Fisher & Gloster, 2005;Monego et al, 2011;Nunes et al, 2014). The subspecies of the M. abscessus complex have undergone taxonomical nomenclature changes, previously classified as three separate subspecies: M. abscessus subsp.…”

Section: Introductionmentioning

confidence: 99%

Investigation of the population structure of Mycobacterium abscessus complex strains using 17-locus variable number tandem repeat typing and the further distinction of Mycobacterium massiliense hsp65 genotypes

Yoshida

Arikawa²,

Tsuyuguchi

et al. 2015

Journal of Medical Microbiology

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Mycobacterium abscessus complex is a significant pathogen in patients with non-cystic fibrosis (non-CF). Nevertheless, there is little description of the genetic diversity of this species. The aims of this study were to investigate the distribution of M. abscessus complex isolated from respiratory specimens by variable number tandem repeat (VNTR) typing. The results of 104 clinical isolates from 104 non-CF patients were compared using PFGE, hsp65 genotypes and clarithromycin susceptibility. The allelic diversity (Hunter-Gaston Discriminatory Index) of the 17 loci examined by VNTR typing was high (0.977). We determined that C28 sequevar erm(41) genotypes and clarithromycin-acquired resistance isolates were scattered in the minimum spanning tree. Intriguingly, VNTR typing and PFGE were highly congruent and revealed that there were clear examples of grouping of isolates from different individuals amongst both M. abscessus and M. massiliense, and showed five clusters of distinct identical isolates. Within these clusters, M. massiliense hsp65 type I formed three different clusters. Although the distribution of M. massiliense hsp65 type II-1 was low (9.3 %), M. massiliense hsp65 type II-1 isolates separated from clusters contained hsp65 type I isolates. Thus, M. massiliense hsp65 genotypes could be discriminated by analysing VNTRs with sufficient genetic distance for intra-species-level discrimination.

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“…bolletii strains. In addition, previous publications have confirmed that outbreak strains, whether harboring the pMAB01 plasmid or not, are fully susceptible to amikacin (AMI) (MIC ϭ 4 g/ml or 8 g/ml) (8)(9)(10).…”

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confidence: 93%

Demonstration of Plasmid-Mediated Drug Resistance in Mycobacterium abscessus

et al. 2014

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b Plasmid-mediated kanamycin resistance was detected in a strain of Mycobacterium abscessus subsp. bolletii responsible for a nationwide epidemic of surgical infections in Brazil. The plasmid did not influence susceptibility to tobramycin, streptomycin, trimethoprim-sulfamethoxazole, clarithromycin, or ciprofloxacin. Plasmid-mediated drug resistance has not been described so far in mycobacteria. P lasmid-mediated drug resistance is uncommon in mycobacteria. A 56,267-bp circular plasmid of the broad-host-range IncP-1␤ subgroup was detected in a strain of Mycobacterium abscessus subsp. bolletii (formerly known as Mycobacterium massiliense [1] and Mycobacterium abscessus subsp. massiliense [2]) that was responsible for a nationwide outbreak of mycobacterial surgical infections in Brazil in 2004Brazil in to 2008. This plasmid, named pMAB01, contains a complete system for conjugative DNA transfer and two genetic load regions carrying antimicrobial resistance genes, including the Tn5393c streptomycin resistance (str) transposon and a typical class 1 integron with an incorporated gene cassette encoding an aminoglycoside 6=-N-acetyltransferase (aac[6=]-Ib), the dihydropteroate synthase type 1 gene (sul1), and a conserved open reading frame (orf5) that carries a GCN5-like N-acetyltransferase (3). We investigated the contribution of this plasmid to antimicrobial drug resistance in M. abscessus and in Escherichia coli.The MICs of streptomycin (STR), kanamycin (KAN), tobramycin (TOB), trimethoprim-sulfamethoxazole (SXT), clarithromycin (CLR), and ciprofloxacin (CIP) were evaluated against the prototype M. abscessus subsp. bolletii outbreak strain INCQS 00594, the INCQS 00594 strain cured of the plasmid after 10 in vitro passages in liquid medium, and two additional outbreak isolates, one harboring the pMAB01 plasmid and one lacking it (Fig. 1A). Two E. coli strains, BL21(DE3) and a spontaneous mutant of E. coli C600 (5) showing nalidixic acid resistance (C600Nal r ), and the same strains harboring the pMAB01 plasmid were tested (Fig. 1B). The experiments performed to introduce pMAB01 into E. coli were detailed in a previous publication (3). In brief, competent E. coli BL21(DE3) isolates were transformed by electroporation with the pMAB01 plasmid purified from the INCQS 00594 isolate. In addition, mating experiments were carried out to confirm the conjugative capacity of the pMAB01 plasmid. Conjugative transfer of pMAB01 from M. abscessus to E. coli BL21(DE3) occurred at a frequency of 5.24 ϫ 10 Ϫ8 transconjugants/recipient and from the E. coli BL21(DE3) transconjugant to another E. coli strain, C600Nal r , at a frequency of 1.14 ϫ 10 Ϫ6 transconjugants/recipient (3). The E. coli C600Nal r strain was used as recipient in mating experiments with two E. coli strains to simplify the screening of E. coli transconjugants on nalidixic acid plates.The MICs were determined by the broth microdilution method according to Clinical and Laboratory Standards Institute documents M24-A2 and M100-S23 (6, 7). M. abscessus subsp. bolletii ...

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Molecular identification and typing of Mycobacterium massiliense isolated from postsurgical infections in Brazil

Abstract: These findings suggest a common source of infection for all patients and reinforce the hypotheses of spread of M. massiliense BRA100 in Brazilian hospital surgical environment in recent years.

Cited by 16 publications

References 25 publications

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

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

Investigation of the population structure of Mycobacterium abscessus complex strains using 17-locus variable number tandem repeat typing and the further distinction of Mycobacterium massiliense hsp65 genotypes

Demonstration of Plasmid-Mediated Drug Resistance in Mycobacterium abscessus

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