Antimicrobial Susceptibility of Standard Strains of Nontuberculous Mycobacteria by Microplate Alamar Blue Assay

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Abstract

In this study, 24 standard nontuberculous mycobacteria (NTM) species strains including 12 slowly growing mycobacteria strains and 12 rapidly growing mycobacteria strains were subjected to drug susceptibility testing using microplate Alamar Blue assay-based 7H9 broth. The most active antimicrobial agents against the 24 NTM strains were streptomycin, amikacin, the fluoroquinolones, and the tetracyclines. Mycobacterium chelonae, Mycobacterium abscessus, Mycobacterium bolletii, and Mycobacterium simiae are resistant to most antimicrobial agents. The susceptibility results of this study from 24 NTM standard strains can be referenced by clinicians before susceptibility testing for clinical isolates is performed or when conditions do not allow for susceptibility testing. The application of broth-based methods is recommended by the Clinical and Laboratory Standards Institute, and the documentation of the susceptibility patterns of standard strains of mycobacteria can improve the international standardization of susceptibility testing methods.

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Most cited references 22

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Mycobacterium abscessus: an emerging rapid-growing potential pathogen.

Björn Petrini (2006)

Mycobacterium abscessus is the most pathogenic and chemotherapy-resistant rapid-growing mycobacterium. It is commonly associated with contaminated traumatic skin wounds and with post-surgical soft tissue infections. It is also one of the mycobacteria that are most often isolated from cystic fibrosis patients. It is essential to differentiate this species from the formerly indistinct "M. chelonae-complex", as chemotherapy is especially difficult in M. abscessussenso strictu. Clarithromycin or azithromycin are the only regular oral antimycobacterial agents with an effect on M. abscessus, and should preferably be supplemented with other drugs since long-term monotherapy may cause resistance. Amikacin is a major parenteral drug against M. abscessus that should also be given in combination with another drug. The recently introduced drug tigecycline may prove to be an important addition to chemotherapy, but has yet to be fully clinically evaluated as an antimycobacterial agent. Surgery can be curative, or at least helpful, in the healing of M. abscessus infection, and if conducted, it should include the removal of all foreign or necrotic material. There is increasing awareness of M. abscessus as an emerging pathogen.

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Comparison of the in vitro activity of the glycylcycline tigecycline (formerly GAR-936) with those of tetracycline, minocycline, and doxycycline against isolates of nontuberculous mycobacteria.

Danny Wilson, Shannon B A Brown, R. Wallace … (2002)

We compared the in vitro activity of the glycylcycline tigecycline (formerly GAR-936) with those of tetracycline, doxycycline, and minocycline by broth microdilution against 76 isolates belonging to seven species of rapidly growing mycobacteria (RGM) and 45 isolates belonging to five species of slowly growing nontuberculous mycobacteria (NTM). By using a resistance breakpoint of >4 micro g/ml for tigecycline and >8 micro g/ml for tetracycline, all RGM were highly susceptible to tigecycline, with inhibition of 50% of isolates at < or =0.12 micro g/ml and inhibition of 90% of isolates at 0.25 micro g/ml for Mycobacterium abscessus and inhibition of both 50 and 90% of isolates at < or =0.12 micro g/ml for M. chelonae and the M. fortuitum group. The MICs of tigecycline were the same for tetracycline-resistant and -susceptible strains, and RGM isolates were 4- to 11-fold more susceptible to tigecycline than to the tetracyclines. In contrast, no slowly growing NTM were susceptible to tigecycline, and isolates of M. marinum and M. kansasii were less susceptible to this agent than to minocycline. This new antimicrobial offers exciting therapeutic potential for the RGM, especially for isolates of the M. chelonae-M. abscessus group, against which the activities of the currently available drugs are limited.

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Evaluation of the GenoType Mycobacterium Assay for identification of mycobacterial species from cultures.

Elvira Richter, Doris Hillemann, Sabine Rüsch-Gerdes (2006)

A new commercially available DNA strip assay (GenoType Mycobacterium CM/AS; Hain Lifescience, Nehren, Germany) was evaluated for the ability to differentiate mycobacterial species. The test is based on a PCR technique targeting a 23S rRNA gene region, followed by reverse hybridization and line probe technology. The GenoType CM is capable of identifying 23, the GenoType AS a further 14, species either alone or in combination with one or more species. Both tests were evaluated with 156 mycobacterial strains composed of 61 validly published species including different subspecies, 6 not validly published species, and 3 strains other than mycobacterial species. All strains were precharacterized by sequencing of the 5' region of the 16S rRNA gene and biochemical tests. In total, results for 151 strains were interpretable. Concordant results were obtained for 137 (92.6%) of 148 mycobacterial strains with the CM assay and 133 (89.9%) of 148 mycobacterial strains with the AS assay, and all three non-Mycobacterium species were identified.

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Author and article information

Contributors

Igor Mokrousov: Role: Editor

Journal

Journal ID (nlm-ta): PLoS One

Journal ID (iso-abbrev): PLoS ONE

Journal ID (publisher-id): plos

Journal ID (pmc): plosone

Title: PLoS ONE

Publisher: Public Library of Science (San Francisco, USA )

ISSN (Electronic): 1932-6203

Publication date Collection: 2013

Publication date (Electronic): 30 December 2013

Volume: 8

Issue: 12

Electronic Location Identifier: e84065

Affiliations

[1 ]National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Beijing, China

[2 ]Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China

[3 ]Pathogenic Biology Institute, University of South China, Hengyang, Hunan, China

[4 ]Key Lab of Laboratory Medicine, Wenzhou Medical College, Wenzhou, Zhejiang, China

St. Petersburg Pasteur Institute, Russian Federation

Author notes

* E-mail: wankanglin@ 123456icdc.cn

Competing Interests: The authors have declared that no competing interests exist.

Conceived and designed the experiments: KW GL. Performed the experiments: GL LLL LW JZ XZ. Analyzed the data: KW GL LLL LW YJ LLZ HL. Contributed reagents/materials/analysis tools: KW GL. Wrote the paper: KW GL.

Article

Publisher ID: PONE-D-13-34257

DOI: 10.1371/journal.pone.0084065

PMC ID: 3875516

PubMed ID: 24386332

SO-VID: 39a593bb-c5bc-4826-8ef2-440c338f799b

License:

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

History

Date received : 13 August 2013

Date accepted : 11 November 2013

Page count

Pages: 6

Funding

This work was financially supported by the projects 2013ZX10003002-001 and 2011ZX10004-001 of the National Key Programme of Mega Infectious Diseases. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Antimicrobial Susceptibility of Standard Strains of Nontuberculous Mycobacteria by Microplate Alamar Blue Assay

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Most cited references 22

Mycobacterium abscessus: an emerging rapid-growing potential pathogen.

Comparison of the in vitro activity of the glycylcycline tigecycline (formerly GAR-936) with those of tetracycline, minocycline, and doxycycline against isolates of nontuberculous mycobacteria.

Evaluation of the GenoType Mycobacterium Assay for identification of mycobacterial species from cultures.

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