Abstract:

Objective: To explore the application value of nanopore sequencing in the diagnosis of tuberculous serous effusion and to evaluate its advantages in rapidly and accurately identifying Mycobacterium tuberculosis and screening for coinfecting pathogens. Methods: From October 2021 to May 2025, patients with suspected tuberculous serous effusion who visited Nanning Fourth People’s Hospital were enrolled. Their effusion samples were collected for nanopore sequencing. Their final clinical diagnoses were used as reference standard for comparing sequencing results with conventional tests (acid-fast staining, TB-DNA PCR, GeneXpert MTB/RIF, and solid mycobacterial culture). Results: Nanopore sequencing achieved significantly better diagnostic performance than any conventional test: sensitivity 73.1% (19/26), specificity 100.0% (27/27), positive predictive value (PPV) 100.0% (19/19), negative predictive value (NPV) 79.4% (27/34), and Kappa 0.734, surpassing those of acid-fast staining (sensitivity 11.5% (3/26), specificity 100.0% (27/27), PPV 3/3, NPV 54.0% (27/50), Kappa 0.117), culture (sensitivity 30.8% (8/26), specificity 100.0% (27/27), PPV 8/8, NPV 60.0% (27/45), Kappa 0.312), TB-DNA PCR (sensitivity 34.6% (9/26), specificity 100.0% (27/27), PPV 9/9, NPV 61.4% (27/44), Kappa 0.350), GeneXpert MTB/RIF (sensitivity 34.6% (9/26), specificity 100.0% (27/27), PPV 9/9, NPV 61.4% (27/44), Kappa 0.350), and TB antibody detection (sensitivity 46.2% (12/26), specificity 81.5% (22/27), PPV 70.6% (12/17), NPV 61.1% (22/36), Kappa 0.278). Moreover, nanopore sequencing revealed that 42.1% (8/19) of tuberculosis cases co-infected with nontuberculous mycobacteria (NTM), viruses, fungi, or bacteria. Conclusion: Nanopore sequencing demonstrated excellent performance in the diagnosis of tuberculous serous effusion, with diagnostic efficacy significantly better than that of traditional pathogen detection methods. It can effectively improve the early diagnosis rate of tuberculosis and offers a new solution for screening coinfections, showing important clinical application value.

Key words: Nanotechnology, Tuberculosis, Pericardial effusion, Pleural effusion, Sensitivity and specificity