Objective　To establish the mFLOS-LAMP method and evaluate its clinical value for rapid detection of Mycobacterium tuberculosis complex (MTBC). Methods February 2022 to December 2023, 42 cases of pulmonary tuberculosis (TB) and 46 patients suffering from other respiratory diseases were selected from the First Affiliated Hospital and the Eighth Affiliated Hospital of Xinjiang Medical University, collect their morning sputum specimens and extract sample DNAUsing the MTBC insertion sequences IS1081 and IS6110 genes as detection targets, a set of specific primers were designed respectively. Based on the design principle of FLOS probes, the ring primers of each set of genes were labeled with corresponding fluorophores, the mFLOS-LAMP method containing dual targets was established, and the detection results of the method were analyzed. Results Using the optimized reaction system, a series of dilutions of genomic DNA of Mycobacterium tuberculosis standard strain H37Rv was used to detect the lower limit of detection of mFLOS-LAMP at 500 fg/μL and the lower limit of detection of mFLOS-PCR at 5 pg/μL and the lower limit of detection of mFLOS-LAMP was 10-fold lower than that of mFLOS-PCR. The mFLOS-LAMP method was non-cross-reactive and highly specific for 24 microbial strains. Clinical specimen testing showed that, using clinical diagnosis results as the reference standard, the sensitivity and specificity of the mFLOS-LAMP method were 92.9% and 100.0%, respectively; the Kappa value was 0.931; and the area under the ROC curves was 0.964. Using the culture method as the gold standard, the sensitivity of the mFLOS-LAMP method was 92.6%. The consistency between the mFLOS-LAMP method and the Xpert MTB/RIF assay was 94.3% (83/88); the Kappa was 0.885, indicating the mFLOS-LAMP method had a high consistency with Xpert MTB/RIF assay. The consistency between the mFLOS-LAMP method and the mFLOS-PCR method was 93.2% (82/88), with a Kappa value of 0.861. Conclusions Compared with the mFLOS-PCR method, the mFLOS-LAMP method has the advantages of rapid detection, high specificity, good sensitivity, high accuracy, and a wider applicability of instrumentation. It can be used as a new type of MTBC detection method.