Mycobacterium tuberculosis, as one of the most serious infectious pathogens, results in millions of deaths and a tremendous economic burden for both individuals and governments. Many potential drugs for the treatment of tuberculosis show excellent activity againstM. tuberculosis in vitro, but lack significant activityin vivowhen administered traditionally by either oral or injection delivery routes. Even though previous researches revealed that inhaled administration method can enhance the efficacy of drugs against mycobacteria, while this evaluating process was complicated and time-consuming by using wild typeM. tuberculosis. Conversely, our research uncovered that the inhalation administration model using strains of autoluminescentM. tuberculosismay provide a more rapid, non-invasive, lung-targeted mid-throughput method, may avoid significant toxic effects on other organs, for evaluating the anti-tuberculosis activity of compounds in mouse models of infection. The live relative light units, quantifying the light emitted by autoluminescentM. tuberculosisH37Rv, can be used to preliminarily evaluate the therapeutic effect of different drug doses, which can significantly shorten the evaluation time. This model offers the ability to assess the efficacy of new lead compounds and TB drugs that can be administered by inhalation.