A rapid and simple modified nitrate reductase assay for testing first and second-line antituberculosis drug susceptibilities in Mycobacterium tuberculosis isolates

In this study, we developed a modified NRA (MONRA) to determine the first and second-line drug susceptibilities of 5 reference ATCC strains and 42 clinical M. tuberculosis isolates. Unlike conventional NRA, which is often performed in solid media or 7H9 broth, the MONRA is performed in a different medium, AYC.2.1 broth, using lyophilized antibiotic tubes to determine drug susceptibility. The MONRA results were compared with BACTEC MGIT 960 method as the reference method for first-line drugs and conventional NRA performed in 7H9 broth for second-line drugs. The agreement between the MONRA and the reference method was determined as 97.62, 100, 97.62, and 100 % for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. When the results were compared with convantional NRA, the agreement was determined as 100 % for all second-line antibiotics including levofloxacin, ofloxacin, and kanamycin. MONRA has the potential to eliminate challenges in implementing drug susceptibility testing in resource-limited settings.