Molecular analysis of Mycobacterium isolates from extrapulmonary specimens obtained from patients in Mexico

Alvarado‐Esquivel, Cosme; García-Corral, Nora; Carrero-Dominguez, David; Enciso-Moreno, José Antonio; Gurrola-Morales, Teodoro; Portillo-Gómez, L; Rossau, R.; Mijs, Wouter

doi:10.1186/1472-6890-9-1

Cited by 24 publications

(24 citation statements)

References 22 publications

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“…M. abscessus is also a leading cause of sporadic and epidemic cases of skin and soft-tissue RGM infections after surgery or following the use of contaminated syringes and needles (21,22,39). Several large outbreaks of skin and soft-tissue infection have been reported following the injection of adrenal cortex extract, mesotherapy, tattooing, and piercing (6,8,15,24,52,58).…”

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Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

et al. 2011

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Mycobacterium abscessus, Mycobacterium bolletii, and Mycobacterium massiliense (Mycobacterium abscessus sensu lato) are closely related species that currently are identified by the sequencing of the rpoB gene. However, recent studies show that rpoB sequencing alone is insufficient to discriminate between these species, and some authors have questioned their current taxonomic classification. We studied here a large collection of M. abscessus (sensu lato) strains by partial rpoB sequencing (752 bp) and multilocus sequence analysis (MLSA). The final MLSA scheme developed was based on the partial sequences of eight housekeeping genes: argH, cya, glpK, gnd, murC, pgm, pta, and purH. The strains studied included the three type strains (M. abscessus CIP 104536 T , M. massiliense CIP 108297 T , and M. bolletii CIP 108541 T ) and 120 isolates recovered between 1997 and 2007 in France, Germany, Switzerland, and Brazil. The rpoB phylogenetic tree confirmed the existence of three main clusters, each comprising the type strain of one species. However, divergence values between the M. massiliense and M. bolletii clusters all were below 3% and between the M. abscessus and M. massiliense clusters were from 2.66 to 3.59%. The tree produced using the concatenated MLSA gene sequences (4,071 bp) also showed three main clusters, each comprising the type strain of one species. The M. abscessus cluster had a bootstrap value of 100% and was mostly compact. Bootstrap values for the M. massiliense and M. bolletii branches were much lower (71 and 61%, respectively), with the M. massiliense cluster having a fuzzy aspect. Mean (range) divergence values were 2.17% (1.13 to 2.58%) between the M. abscessus and M. massiliense clusters, 2.37% (1.5 to 2.85%) between the M. abscessus and M. bolletii clusters, and 2.28% (0.86 to 2.68%) between the M. massiliense and M. bolletii clusters. Adding the rpoB sequence to the MLSA-concatenated sequence (total sequence, 4,823 bp) had little effect on the clustering of strains. We found 10/120 (8.3%) isolates for which the concatenated MLSA gene sequence and rpoB sequence were discordant (e.g., M. massiliense MLSA sequence and M. abscessus rpoB sequence), suggesting the intergroup lateral transfers of rpoB. In conclusion, our study strongly supports the recent proposal that M. abscessus, M. massiliense, and M. bolletii should constitute a single species. Our findings also indicate that there has been a horizontal transfer of rpoB sequences between these subgroups, precluding the use of rpoB sequencing alone for the accurate identification of the two proposed M. abscessus subspecies.

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Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

et al. 2011

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“…The situation of NTM infection throughout the world is the same as in Mexico [13,14], as some NTM were associated with those isolated from reclaimed water like M. avium and M. fortuitum, while M. gordonae was isolated from a water distribution system. Species recovered from a water distribution system were also isolated in the reclaimed water utilised for irrigation.…”

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confidence: 95%

“…M. nonchromogenicum and M. arupense have been isolated from environmental samples and pulmonary disease or synovial tissue and are associated with chronic disease [1,35,36]. The fast growing and opportunistic pathogen M. fortuitum has been isolated from different water samples around the world and it was related to diseases in México by Alvarado-Esquivel [14]; a multicentre study conducted by Esteban et al reported that M. fortuitum was the most common pathogen isolated in lung infection [37]. Thomson [38] reported the increase incidence of M. fortuitum infections per year.…”

Section: Discussionmentioning

confidence: 99%

“…Overall, 12 of the 66 positive cultures were from the Mycobacterium avium complex and 21 of these infections were caused by nonidentified NTM. Recently, Alvarado-Esquivel et al [14] described Mycobacterium species diversity in extra pulmonary-derived specimens in symptomatic patients in Mexico; NTM were found in a considerable number of patients.…”

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confidence: 99%

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Potentially pathogenic nontuberculous mycobacteria found in aquatic systems. Analysis from a reclaimed water and water distribution system in Mexico City

Castillo-Rodal

Mazarí-Hiriart

Lloret-Sánchez

et al. 2011

Eur J Clin Microbiol Infect Dis

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We investigated the presence of nontuberculous mycobacteria (NTM) in three Mexican aquatic systems to evaluate the prevalence with the distribution of NTM species. Key physicochemical parameters of the water samples were determined to find correlations with the species' distributions. We used multilocus sequence analysis (MLSA) based on hsp65, rpoB, and 16S rRNA fragments to determine their taxonomic affiliations. NTM were recovered from water distribution systems and reclaimed water from the Mexico City Metropolitan Area (MCMA). The isolated species were associated with a temperature of 21°C and pH >7.7. The phylogenetic analysis showed that eight of the 14 different NTM strains were unambiguously classifiable: Mycobacterium peregrinum, M. nonchromogenicum (2), M. smegmatis (2), M. fortuitum, M. avium ssp. hominissuis, M. arupense, M. gordonae, and M. chitae. One strain was tentatively identified as M. mantenni/ scrofulaceum and another strain was related to M. porcinum/M. septicum. All NTM species identified in the water distribution system were also detected in the reclaimed water, but some species from the reclaimed water were not found in the water distribution systems. Two of the identified species found in the reclaimed water, M. avium and M. fortuitum, are considered important human opportunistic pathogens.

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“…Disseminated infections caused by M. smegmatis are commonly associated with immunosuppression (18)(19)(20). Alvarado-Esquivel and coworkers have opined that exposure to M. smegmatis might occur through intraoperative contamination or application of adulterated lipid creams to open wounds (21). This organism is also capable of inducing chronic cellulitis with fistula formation, usually as a result of traumatic inoculation of contaminated material (19).…”

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Mycobacterium smegmatis in Skin Biopsy Specimens from Patients with Suppurative Granulomatous Inflammation

Lü

Xia

et al. 2013

J Clin Microbiol

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Formalin-fixed, paraffin-embedded skin biopsy specimens, including 72 suppurative granulomatous inflammation (SGI) and 47 non-SGI controls, were tested for mycobacteria by using a broad-range PCR and a suspension array identification system. Mycobacterium smegmatis was detected in 13 (18.1%) of the SGI skin biopsy specimens, which was significantly more than 2 (4.3%) in the controls (odds ratio, 5.73; 95% confidence interval, 1.21 to 27.06; P ‫؍‬ 0.028).G ranulomatous inflammation of the skin may be broadly classified as infectious or noninfectious, with the presence of a suppurative inflammatory component often associated with the former (1-3). Suppurative granulomatous inflammation (SGI) has been defined as inflammation within the dermis and subcutis consisting of neutrophils, various numbers of eosinophils, and the presence of a histiocytic population of cells (1, 2). Multinucleated giant cells are usually present without obvious foreign material. A diagnosis may be made histologically; however, prior antibiotic exposure may "sterilize" the tissue, leading to a delayed diagnosis and increased morbidity. Tissue culture is the diagnostic gold standard, but when a mycobacterial infection is responsible, cultures often take weeks to grow (4, 5).To date, about one-third of the species of nontuberculous mycobacteria (NTM) described have been associated with human disease (5). Clinical presentations include infections of the skin and soft tissue, lymph nodes, joints, and lungs. Disseminated infections may arise following the contamination of traumatic or surgical wounds with contaminated water or other materials (5, 6). Most species of NTM have been associated with skin and soft tissue infections, but rapidly growing mycobacteria (Mycobacterium fortuitum, M. chelonae, and M. abscessus), M. marinum, and M. ulcerans have been the ones most frequently described. Some of these organisms exhibit geographic restrictions. The timely and accurate diagnosis of cutaneous NTM infections is important in that different species of these organisms have different antibiotic susceptibilities (4, 6, 7). Current laboratory techniques for identifying mycobacteria to the species level include growth in broth, nucleic acid probing, and sequencing, as well as subculture on solid medium for phenotypic testing, each of which is time intensive (4,7,8).We have developed a MycoID assay that incorporates broadrange PCR amplification with suspension array detection to identify 17 mycobacterial complexes, groups, and species in a single reaction with a detection limit of 50 to 500 copies/g of tissue (8). In this study, we have employed the MycoID procedure to detect and identify mycobacterial species in formalin-fixed, paraffin-embedded (FFPE) skin biopsy specimens exhibiting SGI and non-SGI controls.(This study was presented in part at the Association for Molecular Pathology Annual Meeting, Grapevine, TX, 17 to 19 November 2011.)Study materials. The study FFPE tissue specimens consisted of 119 skin biopsy specimens obtained from Beijing Children...

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Molecular analysis of Mycobacterium isolates from extrapulmonary specimens obtained from patients in Mexico

Cited by 24 publications

References 22 publications

Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

Potentially pathogenic nontuberculous mycobacteria found in aquatic systems. Analysis from a reclaimed water and water distribution system in Mexico City

Mycobacterium smegmatis in Skin Biopsy Specimens from Patients with Suppurative Granulomatous Inflammation

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