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[Evaluation of a colorimetric micromethod for determining the minimal inhibitory concentration of antibiotics against Mycobacterium tuberculosis].

2006; National Institutes of Health; Volume: 38; Issue: 3 Linguagem: Inglês

Monica V. Pontino, Beatriz Di Giulio, Carmen Raya Fernández, Belén Imperiale, A Bodon, Nora Morcillo,

Antibiotics Pharmacokinetics and Efficacy

Multidrug-resistant tuberculosis (MDR) caused by strains resistant to both isoniazid and rifampin is now considered a serious sanitary problem worldwide. New technical tools for the early detection of these strains are urgently needed to avoid their spread within the community. We have evaluated a microplate colorimetric-based method to determine the minimal inhibitory concentration (MIC) of first-line antituberculosis drugs by using 3-(4, 5 dimethylthiazolyl 1-2 yl)-2,5 diphenyl tetrazolium bromide as a bacterial growth indicator (MTT) (M-MTT). A total of 603 clinical isolates, 507 from respiratory cases (84.1%) and 96 from non-respiratory cases (15.9%) were processed. The proportion method on a Löwenstein-Jensen medium (PM) with isoniazid (INH), 0.20 microg/ml; streptomycin (SM), 4.00 microg/ml; ethambutol (EMB), 2.00 microg/ml and rifampin (RMP), 40.00 microg/ml, was used as the gold standard. The drugs and the concentration range tested were: INH, 1.00-0.03 microg/ml; SM, 8.00-0.25 microg/ml; EMB, 32.00-1.00 microg/ml and RMP, 2.00-0.06 microg/ml. MIC results were obtained on an average of 8 days (range: 7-12). The cut-off values for each drug, calculated by the ROC curve method, were: INH, 0.25 microg/ml, RMP, 0.50 microg/ml, SM, 4.00 microg/ml and EMB, 4.00 microg/ml. Sensitivity and specificity for RMP were 100 %, while for INH, they were 97.8% and 99.5% respectively. The results obtained suggested that M-MTT is a low cost and easy to set up method that could be applied to MDR clinical diagnosis in developing countries.