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Measuring Minimum Inhibitory Concentrations in Mycobacteria

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Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 465))

Abstract

An agar dilution method for measuring minimum inhibitory concentrations (MICs) of Mycobacterium tuberculosis, based on the method of proportion, is described. Mycobacterium strains are grown on Middlebrook 7H10 (or 7H11) agar medium with twofold serially diluted drug concentrations in order to determine specific inhibitory values. The proportion of bacilli resistant to a given drug is determined by comparing the number of colony-forming units (CFU) on a drug-free control with those growing in the presence of drug within a specific concentration range. The MIC is defined as the lowest drug concentration that inhibits growth of more than 99% of a bacterial population of M. tuberculosis on solid Middlebrook medium within 21 days of incubation at 37°C. The proportion method, the absolute concentration method, and the resistant ratio method have traditionally been used as standard procedures for antimycobacterial drug-susceptibility testing (DST), and reference data are mainly based on these methods. DST concepts and alternative procedures that have been adopted for DST are also briefly discussed.

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Authors and Affiliations

  1. Division of Molecular Biology and Human Genetics, MRC Centre for Molecular and Cellular Biology, DST/NRF Centre of Excellence for Biomedical TB Research, Stellenbosch University, Tygerberg, South Africa

    Frederick A. Sirgel PhD & Ian J.F. Wiid PhD

  2. Division of Molecular Biology and Human Genetics, Stellenbosch University, P.O. Box 19063, Tygerberg, 7505, South Africa

    Paul D. van Helden PhD

Authors
  1. Frederick A. Sirgel PhD
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  2. Ian J.F. Wiid PhD
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  3. Paul D. van Helden PhD
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Corresponding author

Correspondence to Paul D. van Helden PhD .

Editor information

Editors and Affiliations

  1. Queen Mary's School of Med. & Dentistry, Inst. Cell & Molecular Science, Barts and the London, Newark Street 4, London, E1 2AT, United Kingdom

    Tanya Parish

  2. Queen Mary's School of Med. & Dentistry, Inst. Cell & Molecular Science, Barts and the London, Newark Street 4, London, E1 2AT, United Kingdom

    Amanda Claire Brown

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Sirgel, F.A., Wiid, I.J., van Helden, P.D. (2009). Measuring Minimum Inhibitory Concentrations in Mycobacteria. In: Parish, T., Brown, A. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 465. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-207-6_11

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  • DOI: https://doi.org/10.1007/978-1-59745-207-6_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-58829-889-8

  • Online ISBN: 978-1-59745-207-6

  • eBook Packages: Springer Protocols

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