Abstract
Pyrazinamide (PZA) is a key antibiotic in the standard four drug regimen to treat tuberculosis, provided the patient is not PZA resistant. But, determining resistance to PZA phenotypically is difficult, and genotyping the vast array of mutations in the pncA gene is not practical. To circumvent these hurdles, we are developing a robust single-tube method that, at a minimum, can distinguish the wild type drug susceptible pncA gene sequence from all strains that have nonsynonymous genetic alterations (including the variant of M. bovis) by coating them with fluorescent probes in three colors.