ObjectiveTo investigate the value of TSPOT in the diagnosis of tuberculosis（TB） in children. Methods43 children who were admitted to a hospital and confirmed with TB between March 2015 and May 2016 were as TB group, 50 children who were excluded TB as well as healthy children in the outpatient and inpatient departments of this hospital during the same period were as control group, two groups were underwent detection of TSPOT, PPD, MTBAb, and acid fast stain, results were compared, subgroup analysis was conducted on the positive of TSPOT in children with different infection sites and in different age groups. ResultsSensitivity and negative predictive value of TSPOT were significantly higher than the other three methods（P＜0.05），specificity was also higher than that of PPD（P＜0.05）；positive rates of TSPOT in pulmonary tuberculosis and extrapulmonary tuberculosis were 88.9% and 81.3% respectively，difference was not statistically significant（P＞0.05）；positive rates of TSPOT in children ≤5 years and >5 years were 70.6% and 96.2% respectively，difference was statistically significant（P<0.05）. The sensitivity and specificity of combined detection of TSPOT and PPD for diagnosis of TB were 95.3% and 98.0% respectively，which were both higher than the single method（P＜0.05）.ConclusionThe sensitivity and specificity of TSPOT for detecting TB in children are both superior to traditional methods, TSPOT is also suitable for the detection of extrapulmonary tuberculosis, and can be used as the main method for laboratory diagnosis of tuberculosis in children, but it is uncertainty in children under 5 years of age, and should be rationally combined with PPD to improve the diagnosis of TB among children.