e Diagnosis of tuberculous pleurisy remains a challenge in the clinic. In this study, we evaluated the usefulness of a previously developed Mycobacterium tuberculosis antigen-specific gamma interferon enzyme-linked immunospot (ELISPOT) assay in the diagnosis of tuberculous pleurisy by testing a cohort of 352 patients with pleural effusion. We found that M. tuberculosis antigen-specific gamma interferon-producing cells were enriched four to five times in pleural fluid compared with their levels in peripheral blood from patients with tuberuclous pleurisy assayed in parallel. The sensitivity, specificity, positive predictive value, and negative predictive value of the pleural fluid mononuclear cell ELISPOT assay for the diagnosis of tuberculous pleurisy were 95.7%, 100%, 100%, and 81.0%, respectively. In comparison, the sensitivity and specificity of the ELISPOT assay using peripheral blood mononuclear cells were 78.3% and 86.3%, respectively. The sensitivity and specificity of the pleural fluid adenosine deaminase activity test were 55.5% and 86.3%, respectively. These results demonstrate that the M. tuberculosis antigen-specific ELISPOT assay performed on pleural fluid mononuclear cells provides an accurate, rapid diagnosis of tuberculous pleurisy.T uberculosis (TB) is a leading cause of morbidity and mortality throughout the world, with 95% of cases and 97% of all deaths occurring in high-prevalence countries such as China, where the prevalence of active TB is as high as 367/100,000 population (1). Tuberculous pleurisy (TBP) is a common clinical manifestation of active TB disease and may account for up to 50% of all pleural effusions in areas with a high incidence of TB (2). Standard diagnostic assays for TBP, including microbiological examination, adenosine deaminase (ADA) levels, cell infiltrate profile, and certain other biochemical tests, do not provide satisfactory sensitivity and specificity (3). As an example, ADA determination has been recognized as a relatively sensitive diagnostic test for TBP; increased ADA levels are observed in a number of other diseases, such as parapneumonic effusions and noninfectious inflammatory diseases (4). High pleural fluid ADA levels do not necessarily indicate the presence of TBP, especially when pleural fluid potassium levels exceed 5.0 mEq/liter (5).Infection with Mycobacterium tuberculosis elicits a strong antigen-specific gamma interferon (IFN-␥) response from host T cells, which is used as an indicator to diagnose latent M. tuberculosis infection in clinical practice (6, 7). In addition, we and others have applied the M. tuberculosis antigen-specific IFN-␥ enzymelinked immunosorbent spot (ELISPOT) assay to peripheral blood mononuclear cells (PBMC) to diagnose active TB (8, 9). Although the sensitivity of the peripheral blood ELISPOT assay for the diagnosis of active pulmonary TB is significantly higher than those of routine tests such as sputum M. tuberculosis culture, the specificity is not satisfactory, mostly due to its inability to discriminate active TB disease ...