This project will enable high-sensitivity testing of noninvasive oral swab samples for diagnosis of active tuberculosis (TB). Pulmonary TB is usually detected by analysis of sputum, a viscous material from human airways. Sputum collection presents exposure risks and many patients cannot routinely produce sputum for testing, especially in community settings where most TB transmission occurs. In response to the need for alternative, non-invasive sample types, we have demonstrated that Mycobacterium tuberculosis (MTB) DNA can be detected on oral swabs (OS) by using quantitative PCR (qPCR) analysis. In TB OS, the tongue dorsum is gently scraped with a disposable swab. The process is painless and requires just seconds. “Sputum-scarce” people, who can have risk factors for TB but not yet symptoms, can easily be sampled. Self-sampling is straightforward. To further unlock the potential for this approach, the current project will develop and evaluate enhanced swab sampling methods, using new sample processing technologies combined with a unique set of clinical sample resources. Specific Aims are: 1) To develop and assess high-capacity OS testing for pulmonary TB; and 2) To develop and assess protocols for testing serially collected and pooled OS samples for TB screening. TB OS has the potential to simplify TB diagnosis, improve the care of many TB patients, and enable active case-finding strategies that will reduce TB transmission. Thus, this project could help transform the global fight against TB.