This study reports on the modification of surface plasmon resonance (SPR) chips with molybdenum disulfide-molybdenum trioxide (MoS2-MoO3) microflowers to detect the tuberculosis (TB) markers of CFP-10. The MoS2-MoO3 microflowers were prepared by hydrothermal methods with variations in the pH and amount of trisodium citrate (Na3Ct), which were projected to influence the shape and size of microflower particles. The analysis shows that optimum MoS2-MoO3 hybrid microflowers were obtained at neutral pH using 0.5 g Na3Ct. The modified SPR biosensor exhibits a ten times higher response than the bare Au. Moreover, increasing MoS2-MoO3 thickness results in a higher detection response, sensitivity, and a smaller limit of detection (LOD). Using the optimized material composition, the Au/MoS2-MoO3-integrated SPR sensor can demonstrate sensitivity and LOD of 1.005 and 3.45 ng mL-1, respectively. This biosensor also has good selectivity, stability, and reproducibility based on cross-sensitivity characterization with other analytes and repeated measurements on several chips with different storing times and fabrication batch. Therefore, this proposed SPR biosensor possesses high potential to be further developed and applied as a detection technology for CFP-10 in monitoring and diagnosing TB.