BRIEF RESEARCH REPORT article
Front. Cell. Infect. Microbiol.
Sec. Clinical Infectious Diseases
Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1646194
This article is part of the Research TopicExploring Clinical Application Scenarios of Metagenomic Next-Generation Sequencing for Pathogen DiagnosisView all 5 articles
Optimization of decision thresholds for Mycobacterium tuberculosis can effectively improve the performance of mNGS in tuberculosis diagnosis
Provisionally accepted
Yuecui Li1*
Lili Zhang1
Guannan Ma2Chenghang Li1Weiyue Hu1
Ruotong Ren3Yinghui Zang1Dandan Ying1Shuai Qiu1Shuyan Jin1Chunjie Qiu1
Xuefang Cao3*- 1The First People's Hospital of Yongkang, Yongkang, China
- 2Dian Diagnostics Group Co Ltd Key Laboratory of Digital Technology in Medical Diagnostics of Zhejiang Province, Hangzhou, China
- 3MatriDx Biotechnology, Hangzhou, China
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Background: Pulmonary tuberculosis (TB) diagnosis remains challenging due to limitations in traditional methods. This study aimed to optimize the metagenomic next-generation sequencing (mNGS) threshold for Mycobacterium tuberculosis complex (MTBC) detection and evaluate its efficacy compared to standard diagnostic approaches. Methods: A total of 264 bronchoalveolar lavage fluid (BALF) samples were collected from patients with suspected pulmonary TB at Yongkang First People's Hospital between January 2022 and June 2023. After excluding patients with incomplete data, 59 clinically confirmed TB patients and 111 with non-tuberculous conditions were enrolled. mNGS data were analyzed to calculate reads per million (RPM) for MTBC, and thresholds of 0.02, 0.05, and 0.10 RPM were evaluated for diagnostic efficacy using clinical diagnosis as the gold standard. Results: The area under the receiver operating characteristic (ROC) curve (AUC) for mNGS in diagnosing TB at RPM thresholds of ≥0.02, ≥0.05, and ≥0.10 were 4 0.881, 0.873, and 0.814, respectively. The optimal detection threshold was found at RPM ≥ 0.05. Comparative analysis showed mNGS (AUC = 0.873) outperformed routine culture (0.718), PCR (0.741), and Xpert (0.763). Combining mNGS with these methods improved AUC values to 0.837, 0.868, and 0.873, respectively. Conclusion: Optimizing the mNGS threshold to ≥0.05 significantly enhances MTBC detection in pulmonary TB. Combining mNGS with traditional methods further improves diagnostic efficacy, suggesting a potential role for mNGS in clinical TB management.
Keywords: Tuberculosis, Bronchoalveolar Lavage Fluid, metagenomic next-generation sequencing, Threshold optimization, Diagnostic efficacy
Received: 13 Jun 2025; Accepted: 25 Aug 2025.
Copyright: © 2025 Li, Zhang, Ma, Li, Hu, Ren, Zang, Ying, Qiu, Jin, Qiu and Cao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Yuecui Li, The First People's Hospital of Yongkang, Yongkang, China
Xuefang Cao, MatriDx Biotechnology, Hangzhou, China
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