Objective: To analyze the drug resistance characteristics of 231 patients with spinal tuberculosis and to provide reference for clinicians to develop proper treatment regimens for spinal tuberculosis patients. Methods: The Mycobacterium tuberculosis strains and clinical data were collected from 231 culture-positive spinal tuberculosis patients, who were hospitalized in Beijing Chest Hospital from January 2016 to December 2021. Drug susceptibility test was performed for the following 16 drugs using ENCODE microplate methods: streptomycin (Sm), isoniazid (INH), rifampicin (RFP), ethambutol (EMB), rifapentine (Rft), levofloxacin (Lfx), amikacin (Am), capreomycin (Cm), prothionamide (Pto), isoniazid aminosalicylate (Pa), moxifloxacin (Mfx), p-aminosalicylic acid (PAS), clarithromycin (Clr), rifabutin (Rfb), kanamycin (Km) and clofazimine (Cfz). Results: The total drug resistance rate of those spinal tuberculosis patients to at least one of the 16 drugs was 34.63% (80/231), of which the drug resistance rate was significantly higher in previously treated patients (75.44%, 43/57) than in new patients (21.26%, 37/174), and the difference was statistically significant (χ2=55.660, P<0.001). The drug resistance rates of spinal tuberculosis patients to the 16 drugs from high to low were: Sm (24.68%, 57/231)>INH (22.51%, 52/231)>Rft (19.05%, 44/231)>RFP (18.18%, 42/231)>Pa (15.58%, 36/231)>Rfb (13.85%, 32/231)>Lfx (7.79%, 18/231)>PAS (7.36%, 17/231)>Cm (5.63%, 13/231)>Km (4.76%, 11/231)=Cfz (4.76%, 11/231)>Pto (4.33%, 10/231)=Clr (4.33%, 10/231)>EMB (3.90%, 9/231)>Am (3.46%, 8/231)>Mfx (2.60%, 6/231). The poly-drug resistance rate of spinal tuberculosis patients was 11.69% (27/231), of which the poly-drug resistance rate was significantly higher in previously treated patients (19.30%, 11/57) than in new patients (9.20%, 16/174),and the difference was statistically significant (χ2=4.246, P=0.039); The multidrug-resistance rate was 15.58% (36/231), of which the multidrug-resistance rate was significantly higher in previously treated patients (52.63%, 30/57) than in new patients (3.45%, 6/174),and the difference was statistically significant (χ2=46.980, P<0.001). Conclusion: There was serious epidemic of drug resistance in spinal tuberculosis patients. Effective treatment regimens should be developed according to the results of drug susceptibility tests.