High Prevalence of Antimicrobial Resistance in Rapidly Growing Mycobacteria in Taiwan

Yang, Shun-Cheng; Hsueh, Po-Ren; Lai, Hsin‐Chih; Teng, Lee‐Jene; Huang, Li‐Min; Chen, Jong-Min; Wang, Shu-Kuan; Shie, Der-Chuen; Ho, Shen‐Wu; Luh, Kwen‐Tay

doi:10.1128/aac.47.6.1958-1962.2003

Cited by 107 publications

(72 citation statements)

References 30 publications

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“…Another study found that MFLX was active against M. abscessus and that a combination of CAM and MFLX was effective against M. abscessus strains in in vitro models (10). Moreover, some M. abscessus isolates are susceptible to the oral drug linezolid (35,38). However, linezolid was rarely used in our survey to treat M. abscessus species lung disease because of the high cost and moderate to severe side effects in Japan.…”

Section: Discussionmentioning

confidence: 80%

Clinical and Microbiological Differences between Mycobacterium abscessus and Mycobacterium massiliense Lung Diseases

et al. 2012

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In recent years, many novel nontuberculous mycobacterial species have been discovered through genetic analysis. Mycobacterium massiliense and M. bolletii have recently been identified as species separate from M. abscessus. However, little is known regarding their clinical and microbiological differences in Japan. We performed a molecular identification of stored M. abscessus clinical isolates for further identification. We compared clinical characteristics, radiological findings, microbiological findings, and treatment outcomes among patients with M. abscessus and M. massiliense lung diseases. An analysis of 102 previous isolates of M. abscessus identified 72 (71%) M. abscessus, 27 (26%) M. massiliense, and 3 (3%) M. bolletii isolates. Clinical and radiological findings were indistinguishable between the M. abscessus and M. massiliense groups. Forty-two (58%) patients with M. abscessus and 20 (74%) patients with M. massiliense infections received antimicrobial treatment. Both the M. abscessus and M. massiliense groups showed a high level of resistance to all antimicrobials, except for clarithromycin, kanamycin, and amikacin. However, resistance to clarithromycin was more frequently observed in the M. abscessus than in the M. massiliense group (16% and 4%, respectively; P ‫؍‬ 0.145). Moreover, the level of resistance to imipenem was significantly lower in M. abscessus isolates than in M. massiliense isolates (19% and 48%, respectively; P ‫؍‬ 0.007). The proportions of radiological improvement, sputum smear conversion to negativity, and negative culture conversion during the follow-up period were higher in patients with M. massiliense infections than in those with M. abscessus infections. Patients with M. massiliense infections responded more favorably to antimicrobial therapy than those with M. abscessus infections. Mycobacterium species are common causes of pulmonary infections in both humans and animals (14). Although members of the Mycobacterium tuberculosis complex cause the majority of pulmonary infections worldwide, many nontuberculous mycobacteria (NTM) can cause similar infections (13,20). In recent years, many novel NTM species have been discovered through the increased application of genetic investigation tools; detailed genetic characterizations have helped define new taxonomic groupings (17,29). Recently, two new M. abscessus-related species, M. massiliense and M. bolletii, were identified, which were previously grouped with M. abscessus (1, 3). The rate of isolation of these two species has been increasing in Japan. However, very little is known about the natural epidemiology and pathogenicity of M. massiliense and M. bolletii outside outbreak situations. One report found that the ratio of M. abscessus to all NTM is much higher in South Korea (19) than in other countries, including Japan.Here, we aimed to evaluate the epidemiology, clinical and radiological spectrum, treatments, drug susceptibility, and outcome of M. abscessus and M. massiliense lung diseases during therapy in Japan. MATERIALS AND ME...

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Section: Discussionmentioning

confidence: 80%

Clinical and Microbiological Differences between Mycobacterium abscessus and Mycobacterium massiliense Lung Diseases

et al. 2012

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“…M. abscessus is also recovered from respiratory specimens of patients with cystic fibrosis (1,4). The differences in antimicrobial susceptibilities between M. abscessus and M. chelonae demand an easy differentiation of these two closely related species (14). Restriction analysis of an hsp65 gene fragment has been used to this end for over 10 years (7,12,13).…”

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LightCycler-Based Differentiation of Mycobacterium abscessus and Mycobacterium chelonae

et al. 2004

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In this study we introduce a rapid procedure to identify Mycobacterium abscessus (types I and II) and M. chelonae using LightCycler-based analysis of the hsp65 gene. Results from 36 clinical strains were compared with hsp65 gene restriction analysis and biochemical profiles of bacilli. As all three methods yielded identical results for each isolate, this procedure offers an excellent alternative to previously established nucleic acid amplification-based techniques for the diagnosis of mycobacterial diseases.Among frequently isolated clinical strains of rapidly growing mycobacteria are Mycobacterium abscessus and Mycobacterium chelonae (2). Patients typically present with local abscesses, lymphadenitis, and orbital or pulmonary infections (10, 11). M. abscessus is also recovered from respiratory specimens of patients with cystic fibrosis (1, 4). The differences in antimicrobial susceptibilities between M. abscessus and M. chelonae demand an easy differentiation of these two closely related species (14). Restriction analysis of an hsp65 gene fragment has been used to this end for over 10 years (7,12,13). However, this method requires subsequent gel analysis of amplified DNA. Likewise, interpretation of fragment sizes is sometimes difficult because of the similarity of restriction patterns. Alternative diagnostic procedures include glycan analysis of the bacterial capsule using high-performance liquid chromatography, which is highly reliable but can be performed only in specialized laboratories (13). In this study, we developed a simple and rapid method based on LightCycler technology to distinguish M. abscessus and M. chelonae. We compared the biochemical profiles, the restriction patterns of the hsp65 gene, and partial sequences of the hsp65 gene of 36 clinical strains, including M. abscessus strain ATCC 19977 and M. chelonae strain ATCC 35752 as reference strains. All strains had previously been identified as belonging to the M. chelonae-M. abscessus group by 16S rRNA gene analysis as described previously (5).Biochemical profiling included testing for growth on 5% sodium chloride and growth on citrate as the sole carbon source as suggested by Yakrus and colleagues (13). Briefly, bacteria were precultured in liquid medium using mycobacterial growth indicator tubes (MGIT) (Becton Dickinson, Oxford, United Kingdom). Positive MGIT cultures were subcultured with 0.1 ml of a 1:10 dilution on Lowenstein-Jensen selective medium (L-J) containing 5% NaCl (Remel, Lenexa, Kans.). Utilization of sodium citrate was determined as described by Silcox et al. (8). Briefly, a basal medium was prepared by dissolving 2.4 g of (NH 4 ) 2 SO 4 (Merck, Darmstadt, Germany), 0.5 g of KH 2 PO 4 (Merck), and 0.5 g of MgSO 4 · 7H 2 O (Merck) in 950 ml of distilled water; the pH was adjusted to 7.0, and after adding 20 g of Noble agar (Difco Laboratories, Detroit, Mich.), the mixture was autoclaved. Sodium citrate (5.6 g) (J. T. Baker, Deventer, The Netherlands) was dissolved in 50 ml of distilled water, filter sterilized, and added aseptical...

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“…This limitation has no clinical relevance currently for the diagnosis of infection by M. abscessus except for the possibility of epidemiological analysis in clinical infection. The most important benefit of the ITS PCR assay is the ability to differentiate between M. chelonae and M. abscessus, since infections with these organisms require different treatment regimens (16,17).…”

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Use of the MGB Eclipse System and SmartCycler PCR for Differentiation of Mycobacterium chelonae and M. abscessus

et al. 2005

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Although accurate in the identification of Mycobacterium species, partial 16S rRNA gene sequencing does not distinguish Mycobacterium chelonae from M. abscessus. Thus, we designed a SmartCycler PCR assay targeting the 16S-to-23S internal transcribed spacer (ITS) region with use of MGB Eclipse probes to distinguish each species. Comparison with PCR-restriction enzyme analysis of a 441-bp fragment of the hsp65 gene resulted in 100% correlation with 25 isolates of M. chelonae and 25 isolates of M. abscessus. ITS PCR performed on 90 consecutive isolates identified by partial 16S rRNA gene sequencing (26 isolates of the M. chelonae-M. abscessus complex and 64 remaining isolates, including Mycobacterium species, Nocardia species, and other aerobic actinomycetes) showed 100% specificity and sensitivity. The ITS PCR assay is accurate and specific, easy to perform, and a good supplemental test when using partial 16S rRNA gene sequencing to identify M. chelonae and M. abscessus.The use of partial (5Ј end, approximately 500 bp) 16S rRNA gene sequencing is an accurate method for the identification of Mycobacterium species from cultured isolates (1,3,5,8,10,14). This method, however, does not differentiate Mycobacterium chelonae from M. abscessus. Conventional tests that have been used to separate these two species include growth on 5% sodium chloride and use of citrate medium as a carbon source (16). Limitations of the conventional tests include difficulty interpreting results (2) and prolonged turnaround time, requiring up to 3 weeks of incubation. Restriction enzyme digestion and analysis of an amplified fragment (441 bp) of the hsp65 gene can distinguish M. chelonae from M. abscessus but requires gel analysis and interpretation of fragment sizes and is sometimes difficult because of the similarity of restriction patterns (11). The objective of this study was to develop a simple and rapid method, using real-time PCR with the SmartCycler (Cepheid, Sunnyvale, CA), that would differentiate between M. abscessus and M. chelonae.To initiate the design of primers and probes, sequences of the 16S-to-23S internal transcribed spacer (ITS) region of several Mycobacterium species were either determined by ARUP laboratories or retrieved

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High Prevalence of Antimicrobial Resistance in Rapidly Growing Mycobacteria in Taiwan

Cited by 107 publications

References 30 publications

Clinical and Microbiological Differences between Mycobacterium abscessus and Mycobacterium massiliense Lung Diseases

Clinical and Microbiological Differences between Mycobacterium abscessus and Mycobacterium massiliense Lung Diseases

LightCycler-Based Differentiation of Mycobacterium abscessus and Mycobacterium chelonae

Use of the MGB Eclipse System and SmartCycler PCR for Differentiation of Mycobacterium chelonae and M. abscessus

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