Abstract
The aim of this study was to develop and evaluate two colorimetric nitrate reductase-based antibiotic susceptibility (CONRAS) tests, the CONRAS-liquid test and the CONRAS-LJ test, to enable susceptibility testing of Mycobacterium tuberculosis to pyrazinamide. To enhance the growth of M. tuberculosis in 7H9 broth with acid pH (6.0), the effect of three potential growth-promoting substances (reconstitution fluid, fastidious organism supplement and epidermal growth factor) was evaluated. Seventy-five M. tuberculosis strains were tested for susceptibility to pyrazinamide in the CONRAS-liquid test performed in Middlebrook 7H9 broth, and 77 M. tuberculosis strains were tested for susceptibility to pyrazinamide in the CONRAS-LJ test performed on Löwenstein-Jensen medium. The BACTEC 460TB system, sequencing of the pncA gene and Wayne's assay were used as reference tests. Growth of 10 M. tuberculosis strains in conventional 7H9 broth and in acid 7H9 broth with and without growth-promoting substances added was evaluated by the nitrate reductase assay. By using the BACTEC 460TB system as the reference test, the sensitivity and specificity of the CONRAS-liquid test (100 mg/L pyrazinamide) tested on 75 M. tuberculosis strains were 87.5% and...Continue Reading
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