〔Abstract〕 Objective To explore the application value of Xpert MTB/RIF test probes in rifampicin-resistant patients. Methods A total of 548 specimens collected from the third people's Hospital of Shenzhen were analyzed, including sputum, bronchoalveolar lavage fluid, pleural and ascites, cerebrospinal fluid, urine and so on (2018.6 to 2021.5). The Xpert MTB/RIF assay and absolute concentration susceptibility test were performed simultaneously. Through the analysis of the two results, the relationship between the probes of Xpert MTB/RIF assay and the resistance of rifampin and rifampin was obtained. Results The Xpert test result is positive for any probe, which indicates that the comprehensive coincidence rate of the RFB cross-resistance test is 51.64%. The strains with positive A, C, D+E probes in the Xpert test indicate that they are resistant to RFP, but are all sensitive to RFB. Other probes are of little significance as an indicator of drug sensitivity. Using the absolute concentration method as the gold standard, the Xpert detection Results for the resistance of each probe to RFB have different sensitivity and specificity. Conclusion The Xpert test Results are positive for some probes, which can be used as an indicator for RFP-resistant patients to select RFB replacement.