Literature DB >> 23750367

Mycobacterium tuberculosis Beijing type mutation frequency--author's response.

Jurriaan E M de Steenwinkel, Dick van Soolingen, Irma A J M Bakker-Woudenberg.   

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Year:  2013        PMID: 23750367      PMCID: PMC3647678          DOI: 10.3201/eid1903.121849

Source DB:  PubMed          Journal:  Emerg Infect Dis        ISSN: 1080-6040            Impact factor:   6.883


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In Response: We explain the differing frequencies of mutation to rifampin resistance mentioned by Werngren (). First, the strains of Mycobacterium tuberculosis that we tested differed from those previously tested (). Second, we used different rifampin concentrations in subculture plates. For Beijing strain 2002–1585, Bergval et al. () found a mutation frequency of 4–24 × 10−8 at a subculture concentration of 8 mg/L, whereas we found a mutation frequency of 3–4 × 10−3 at a subculture concentration of 1 mg/L and a lower mutation frequency at 2 mg/L. Thus, the concentration of drugs in subculture plates is crucial to mutation frequency assays. Absent a subculture concentration standard, we applied rifampin at 1 mg/L () because bacteria growing at this concentration are considered resistant to rifampin. Our mutation frequency and time-kill kinetics assay results are not contradictory because in the time-kill kinetics assays, the subculture rifampin concentration was 4 mg/L. We performed no classical fluctuation assays. We compared the Beijing genotype with the East African/Indian genotype to learn how M. tuberculosis strains differed in their capacity to withstand antituberculosis drug treatment. For reference strain H37Rv, mutation frequency was 1.5 × 10−6, higher than that found with higher subculture concentrations. With regard to the 3 other issues, our drug-susceptibility testing of mutants showed a stable rifampin-resistant phenotype. We agree that these bacteria might represent preexisting mutants selected during drug exposure in a certain drug concentration window. By using different concentrations in subculture plates in our mutation frequency assay, we detected such preexisting mutants. Heteroresistance probably does not explain our observations because in our time-kill kinetics experiments, the whole mycobacterial population decreased over time in a drug concentration-dependent way, and regrowth of a drug-resistant subpopulation was not observed. By not sticking to the fixed test conditions as used in the classical drug-susceptibility assays, research leads to highly interesting findings. One can conclude that serendipity flourishes with variation.
  3 in total

1.  Drug-susceptible Mycobacterium tuberculosis Beijing genotype does not develop mutation-conferred resistance to rifampin at an elevated rate.

Authors:  Jim Werngren; Sven E Hoffner
Journal:  J Clin Microbiol       Date:  2003-04       Impact factor: 5.948

2.  Resistant mutants of Mycobacterium tuberculosis selected in vitro do not reflect the in vivo mechanism of isoniazid resistance.

Authors:  Indra L Bergval; Anja R J Schuitema; Paul R Klatser; Richard M Anthony
Journal:  J Antimicrob Chemother       Date:  2009-07-04       Impact factor: 5.790

3.  Mycobacterium tuberculosis Beijing type mutation frequency.

Authors:  Jim Werngren
Journal:  Emerg Infect Dis       Date:  2013-03       Impact factor: 6.883

  3 in total

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