Rapid detection of mycobacterium tuberculosis complex in sputum specimens using MinION sequencing:sensitivi-ty and specificity evaluation
XIA Liang-hua
YU Sheng-ming
XIE Ling
YAN Liang
CHEN Kai-sen
XIE Rong-hui
Abstract：Objective To evaluate the sensitivity,specificity,and diagnostic performance of MinION sequencing technology for rapid detection of Mycobacterium tuberculosis complex(MTBC)in sputum specimens,compared with acid-fast staining microscopy and quantitative PCR(qPCR),and to assess its stability and reproducibility for potential clinical application.Methods Sputum samples were spiked with varying concentrations of M.tuberculosis H37Rv stand-ard strain to simulate clinical infection and analyzed using acid-fast staining,qPCR,and MinION sequencing to deter-mine sensitivity.To assess specificity,sputum samples were spiked with different non-tuberculous mycobacteria species,DNA was extracted,and analyzed via MinION sequencing.The minimum detection limit,reproducibility,and cross-re-activity were evaluated.Results MinION sequencing detected MTBC at a minimum concentration of 104 cfu/mL with a specificity of 100％.However,reproducibility was suboptimal,and detection thresholds were relatively high compared to conventional methods.Conclusion MinION sequencing enables direct detection of MTBC in sputum samples with high specificity but exhibits limited reproducibility and sensitivity at lower bacterial loads.Further validation and data are re-quired before routine clinical implementation.
Keywords：MinION sequencingdetection of Mycobacterium tuberculosis complexsensitivityspecificity
Publication Date：2026-01-15
Online Publishing Date：2026-03-04(First online date of this platform, not the publication date of the document)
Pages：6( 23-28 )