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Mycobacteriology and Aerobic Actinomycetes

Validation of the Use of Middlebrook 7H10 Agar, BACTEC MGIT 960, and BACTEC 460 12B Media for Testing the Susceptibility of Mycobacterium tuberculosis to Levofloxacin

Cynthia A. Sanders, Rachel R. Nieda, Edward P. Desmond
Cynthia A. Sanders
California Department of Health Services, Richmond, California
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Rachel R. Nieda
California Department of Health Services, Richmond, California
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Edward P. Desmond
California Department of Health Services, Richmond, California
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  • For correspondence: edesmond@dhs.ca.gov
DOI: 10.1128/JCM.42.11.5225-5228.2004
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    FIG. 1.

    LVX MIC results for susceptible and resistant clinical isolates of M. tuberculosis. LVX MIC results for BACTEC 460, BACTEC MGIT 960, and AP methods are shown for 32 pan-susceptible clinical isolates of M. tuberculosis and 14 M. tuberculosis isolates that either were clinically resistant to LVX or had known DNA gyrase resistance mutations.

Tables

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  • TABLE 1.

    Drug stability study of BACTECa

    LVX concn (μg/ml)TreatmentReading no.
    12345Avg
    2.0Fresh394443414442.2
    4°C414145354641.6
    37°C403742404140.0
    1.0Fresh9081757810385.4
    4°C706186709476.2
    37°C1316346575871.0
    • ↵ a Results of the drug stability studies in the BACTEC system for LVX at two concentrations in freshly prepared drug-containing medium, medium stored at 4°C for 14 days, and medium stored at 37°C for 14 days inoculated with M. tuberculosis strain H37Rv. GI readings in units at the time the drug-free control reached a GI of ≥30 (5 days) for the various treatments are shown.

  • TABLE 2.

    Drug stability study with BACTEC MGIT 960 systema

    LVX concn (μg/ml)TreatmentNo. of days to positivity for reading no.
    12345Avg
    0.5Fresh16.019.818.217.221.118.5
    4°C24.523.416.225.923.522.7
    37°C24.521.116.827.918.821.8
    0.25Fresh5.35.45.84.66.15.4
    4°C6.05.35.56.14.35.4
    37°C4.74.95.24.84.84.9
    • ↵ a Results of the drug stability studies in the BACTEC MGIT 960 system for LVX at two subinhibitory concentrations in freshly prepared drug-containing medium, medium stored at 4°C for 14 days, and medium stored at 37°C for 14 days inoculated with M. tuberculosis strain H37Rv. The number of days to positivity indicates the number of days to achieve detectable growth with the BACTEC MGIT 960 instrument. A control with no drug was also tested and became positive in 3.3 to 3.5 days.

  • TABLE 3.

    Results of critical concentration testing of three methodsa

    MethodLVX test concn (μg/ml)No. of isolates with susceptible results/No. susceptibleNo. of isolates with resistant results/No. resistant
    BACTEC 46028/9b14/14
    BACTEC MGIT 96029/914/14
    AP123/24c13/14d
    • ↵ a Results of LVX critical concentration testing for the BACTEC 460, BACTEC MGIT 960 and AP methods. The critical test concentrations of LVX were chosen for each method based on results of testing clinical isolates of M. tuberculosis with known LVX susceptibilities. The selected critical test concentrations were then tested against a subset of LVX-susceptible M. tuberculosis isolates as well as isolates that were either clinically resistant to LVX or had known gyrA mutations associated with quinolone resistance.

    • ↵ b One isolate failed to grow in 12B medium.

    • ↵ c One 7H10 plate was unevaluable due to contamination.

    • ↵ d One isolate tested borderline resistant (1 to 10%). One isolate failed to grow on 7H10 medium.

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Validation of the Use of Middlebrook 7H10 Agar, BACTEC MGIT 960, and BACTEC 460 12B Media for Testing the Susceptibility of Mycobacterium tuberculosis to Levofloxacin
Cynthia A. Sanders, Rachel R. Nieda, Edward P. Desmond
Journal of Clinical Microbiology Nov 2004, 42 (11) 5225-5228; DOI: 10.1128/JCM.42.11.5225-5228.2004

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Validation of the Use of Middlebrook 7H10 Agar, BACTEC MGIT 960, and BACTEC 460 12B Media for Testing the Susceptibility of Mycobacterium tuberculosis to Levofloxacin
Cynthia A. Sanders, Rachel R. Nieda, Edward P. Desmond
Journal of Clinical Microbiology Nov 2004, 42 (11) 5225-5228; DOI: 10.1128/JCM.42.11.5225-5228.2004
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KEYWORDS

Anti-Bacterial Agents
levofloxacin
Mycobacterium tuberculosis
Ofloxacin

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