Background: Xpert MTB/RIF assay is a rapid automated molecular test with excellent reported sensitivity, specificity for diagnosis of pulmonary and extrapulmonary Mycobacterium tuberculosis (MTB) infections. However, the clinical utility and accuracy in STB is not well established. A study was conducted to report on the sensitivity, specificity and clinical utility of the Xpert MTB/RIF assay in spinal tuberculosis (STB). Methods: A retrospective review of medical records was performed for 136 patients that underwent spinal biopsy for suspected spondylodiscitis. Reports for acid fast bacilli (AFB) smear, gram stain, pyogenic culture, MTB culture, histopathology, Xpert MTB/RIF assay, and drug sensitivity testing were reviewed. 'Reference standard for diagnosis of STB' was based on positive histopathology and/or MTB culture evidence and was considered as MTB positive. Any samples returning a positive pyogenic or fungal culture were considered as MTB negative. The sensitivity, specificity for Xpert MTB/RIF was assessed against the reference standard. Results: A total of 125 patients were considered for final analysis, 86 patients met the criteria for 'Reference standard for diagnosis of TB spine' (MTB positive). This includes nine patients that were MGIT culture only positive; 45 that were histopathology only positive and 32 were both culture and histopathology positive. There were 39 culture proven (pyogenic-37 and fungal-2) patients included in MTB negative group. The 86 MTB positive patients, included 53 (61.6%) tissue samples and 33 (38.4%) pus samples. The overall analysis showed a 65.1% sensitivity, 100% specificity, 100% PPV and 56.5% NPV for the Xpert MTB/RIF. Conclusions: Gene Xpert MTB/RIF showed excellent specificity and was accurate in the identification of drug resistance. The sensitivity was 65% and sampling techniques using pus samples rather than tissue samples could be a possible reason for lower sensitivity.