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Antimicrobial susceptibility of rapidly growing mycobacteria using the rapid colorimetric method

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Abstract

Drug susceptibility testing (DST) of rapidly growing mycobacteria (RGM) are recommended for guiding the antimicrobial therapy. We have evaluated the use of resazurin in Mueller–Hinton medium (MHR) for MIC determination of RGM and compared the results with those obtained with the reference standard broth microdilution in Mueller–Hinton (MH) and with the resazurin microtiter assay (REMA) in 7H9 broth. The MIC of eight drugs: amikacin (AMI), cefoxitin (FOX), ciprofloxacin (CIP), clarithromycin (CLA), doxycycline (DOX), linezolid (LZD), moxifloxacin (MXF) and trimethoprim-sulfamethoxazole (TMP-SMX) were evaluated against 76 RGM (18 species) using three methods (MH, MHR, and REMA) in a 96-well plate format incubated at 37 °C over 3–5 days. Results obtained in the MH plates were interpreted by the appearance of turbidity at the bottom of the well before adding the resazurin. MHR and 7H9-REMA plates were read by visual observation for a change in color from blue to pink. The majority of results were obtained at day 5 for MH and 1 day after for MHR and 7H9-REMA. However, the preliminary experiment on time to positivity results using the reference strain showed that the resazurin can be added to the MH at day 2 to produce the results at day 3, but future studies with large sets of strains are required to confirm this suggestion. A high level of agreement (kappa 1.000–0.884) was obtained between the MH and the MHR. Comparison of results obtained with 7H9-REMA, on the other hand, revealed several discrepancies and a lower level of agreement (kappa 1.000–0.111). The majority of the strains were resistant to DOX and TMP-SMX, and the most active antimicrobials for RGM were AMI and FOX. In the present study, MHR represented an excellent alternative for MIC determination of RGM. The results could be read reliably, more easily, and more quickly than with the classical MH method.

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Acknowledgements

We thank Dr. Edward Moore for kindly providing us with CCUG type and reference strains. We acknowledge the “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior” (CAPES, Brazil) for the financial support to Ivy Bastos Ramis (PDSE-7675/13-5).

Conflict of interest

The authors declare that they have no conflicts of interest.

Compliance with ethical standards

We confirm that no human participants or animals were included in this study. Consequently, no informed consent was needed.

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Authors and Affiliations

  1. Centro de Desenvolvimento Tecnológico (CDTec), Universidade Federal de Pelotas, Pelotas, Brazil

    I. B. Ramis & P. E. A. da Silva

  2. Laboratory of Microbiology, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium

    I. B. Ramis, M. Cnockaert, J. C. Palomino, P. Vandamme & A. Martin

  3. Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil

    A. von Groll, C. L. Nogueira & S. C. Leão

  4. Pôle de Microbiologie, Institut de Recherche Expérimentale et Clinique, UC Louvain, Brussels, Belgium

    E. Andre & A. Simon

  5. Laboratório de Micobacteriologia, Faculdade de Medicina, Universidade Federal do Rio Grande, Rio Grande, Brazil

    P. E. A. da Silva

Authors
  1. I. B. Ramis
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  2. M. Cnockaert
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  3. A. von Groll
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  4. C. L. Nogueira
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  5. S. C. Leão
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  6. E. Andre
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  7. A. Simon
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  8. J. C. Palomino
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  9. P. E. A. da Silva
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  10. P. Vandamme
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  11. A. Martin
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Correspondence to I. B. Ramis.

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Ramis, I.B., Cnockaert, M., von Groll, A. et al. Antimicrobial susceptibility of rapidly growing mycobacteria using the rapid colorimetric method. Eur J Clin Microbiol Infect Dis 34, 1403–1413 (2015). https://doi.org/10.1007/s10096-015-2365-2

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  • DOI: https://doi.org/10.1007/s10096-015-2365-2

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