Abstract: Objective  To investigate the clinical value of mycobacterial DNA microarray technology for diagnosis of childhood tuberculosis.   Methods  120 clinical specimens were collected from hospitalized child patients. Acid-fast staining, mycobacterial culture and DNA microarray assays were performed using these clinical specimens. The results of DNA microarray assays were compared with the results of acid-fast staining and mycobacterial culture.   Results  The sensitivity of DNA microarray assays for specimens from children with tuberculosis was 24.3% (17/70), of acid-fast staining 17.1% (12/70), of mycobacterial culture 20.0% (14/70), and the specificity of the three methods was all 100.0% (50/50). The difference between results of DNA microarray assays and that of acid-fast staining or mycobacterial culture was not significant.   Conclusion  DNA microarray assay has reference value for the diagnosis of childhood tuberculosis. It provides a new way for the diagnosis of childhood tuberculosis.