The clinical data of 801 human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) patients with suspected tuberculosis were collected to explore the diagnosis value of Xpert Mycobacterium tuberculosis/rifampicin (Xpert MTB/RIF) for tuberculosis in HIV/AIDS patients admitted to the Department of Infection and Immunology of Shanghai Public Health Clinical Center affiliated to Fudan University between January 1, 2018 and December 31, 2020. Among 801 HIV/AIDS patients with suspected tuberculosis, 657 patients received tuberculosis screening tests including Xpert MTB/RIF, peripheral blood tuberculosis T cell spot test (T-SPOT.TB), smear microscopy and BACTEC MGIT 960 liquid culture. When the results of liquid culture and Mycobacterium identification and typing were set as the diagnostic criterion for tuberculosis, the sensitivities of Xpert MTB/RIF, T-SPOT.TB, smear microscopy in tuberculosis diagnosis among HIV/AIDS patients (including pulmonary tuberculosis and extra-pulmonary tuberculosis) were 72.8%, 55.4%, 69.6%, respectively, the specificities were 96.8%, 90.3%, 84.4%, respectively, and Kappa values were 0.719 (P<0.01), 0.430 (P<0.01), 0.424 (P<0.01), respectively. The sensitivity and specificity of Xpert MTB/RIF in the diagnosis of pulmonary tuberculosis were 66.7% and 96.0%, respectively. Among sputum smear-positive and sputum smear-negative tuberculosis patients, the sensitivity of Xpert MTB/RIF was 77.4% and 35.2%, respectively, and the specificity was 87.7% and 97.8%, respectively. The sensitivity and specificity of Xpert MTB/RIF in the diagnosis of extra-pulmonary tuberculosis were 63.3% and 95.2%, respectively. The results indicate that Xpert MTB/RIF has high sensitivity and specificity in the diagnosis of tuberculosis in HIV/AIDS patients, and the sensitivity in the diagnosis of pulmonary tuberculosis is higher than that of extra-pulmonary tuberculosis. It is recommended that Xpert MTB/RIF should be used as the preferred testing technique for suspected tuberculosis in HIV/AIDS patients.