Abstract: OBJECTIVE To explore the efficiency of T-cell spot test for tuberculosis (T-SPOT.TB) combined with cytokines in differential diagnosis of active tuberculosis (ATB) and analyze the influencing factors. METHODS A total of 240 patients who were treated in Hebei Chest Hospital from Jan. 2019 to Oct. 2023 were recruited as the research subjects and were divided into the ATB group with 123 cases and the non-ATB group with 117 cases. The efficiency of T-SPOT.TB combined with cytokines in differential diagnosis of ATB was analyzed by the area under the receiver operating characteristic (ROC) curve, the consistency was analyzed by means of Kappa consistency check. Multivariate logistic regression analysis was performed for the influencing factors for the diagnostic accuracy of T-SPOT.TB. RESULTS There were significant differences in history of contact with tuberculosis, total protein in pleural effusion, pulmonary bacteria infection, positive test for tuberculin (PPD), positive test for tuberculosis-antibody (TB-Ab) , positive T-SPOT.TB for pleural effusion, diameter of cavity, pleural thickening, pleural adhesion, erythema nodosum and richness of blood supply between the two groups of patients (P<0.05). The levels of interleukin (IL)-6, IL-8 and interferon-γ (IFN-γ) of the ATB group were (43.98±21.32)ng/L, (27.95±12.26)ng/L and (40.75±19.11)ng/L, respectively, higher than those of the non-ATB group (P<0.05). The receiver operating characteristic (ROC) curve analysis showed that the AUC of the joint detection of cytokines IFN-γ, IL-6, IL-8 and T-SPOT.TB counts was greater than that of the single detection of the above indexes (P<0.05); the pulmonary bacterial infection was an influencing factor for the accuracy of T-SPOT.TB in diagnosis (P=0.018,OR=5.703,95%CI:1.358 to 23.953). CONCLUSION T-SPOT.TB combined with IFN-γ, IL-6 and IL-8 shows favorable efficiency in diagnosis of ATB and is worthy to be promoted in the hospital.