Gou Boya, Zhang Chuyao, Song Zhe, Huang Zhen, Lu Shuihua
National Clinical Research Center for Infectious Diseases, Shenzhen Third People's Hospital, Shenzhen 518112, China.
Department of Biochemistry and Molecular Biology, Tulane University School of Medicine, New Orleans, LA 70112, USA; Center for Cellular and Molecular Diagnostics, Tulane University School of Medicine, New Orleans, LA 70112, USA.
Clin Chim Acta. 2025 Sep 1;577:120469. doi: 10.1016/j.cca.2025.120469. Epub 2025 Jul 7.
Targeted next-generation sequencing (tNGS) is an adjunct tool endorsed by the World Health Organization (WHO) for tuberculosis diagnosis. The study aimed to investigate the diagnostic performance of tNGS across different clinical scenarios (populations, sample and tuberculosis types, and platforms), which has not been comprehensively evaluated.
A systematic literature search was conducted in PubMed, MEDLINE, Web of Science, Wanfang Data, VIP, and CNKI to identify studies published in English and Chinese from January 1, 2005, to October 14, 2024, that evaluated tNGS for tuberculosis diagnosis using reference strains and original samples or isolates from patients with presumptive or confirmed tuberculosis. Included studies were assessed using the Quality Assessment of Diagnostic Accuracy Studies-2 tool (QUADAS-2), and diagnostic accuracy was calculated against microbiological reference standard and composite reference standard. The primary outcomes were the pooled sensitivity and specificity. This study is registered with PROSPERO (CRD42024560190).
Out of 3,383 records screened, 16 studies comprising 2,565 participants were included in the meta-analysis. Overall, tNGS demonstrated a pooled sensitivity of 0.86 (95 % CI, 0.77-0.91) and specificity of 0.95 (0.86-0.99). The positive and negative likelihood ratios were 18.24 (5.61-59.26) and 0.15 (0.09-0.26). The diagnostic odds ratio and the area under the summary receiver operating characteristic curve was 120.88 (26.93-542.57) and 0.95 (0.93-0.97), respectively. The diagnostic performance of tNGS varied substantially depending on sample type and sequencing platform, where the highest sensitivity was observed with bronchoalveolar lavage fluid 0.91 (0.75-0.98) and the target nanopore sequencing platform 0.85 (0.80-0.89). Furthermore, tNGS demonstrated a significantly higher detection rate compared to GeneXpert MTB/RIF (odds ratio: 1.78 (1.39-2.29); p = 0.01).
tNGS demonstrates high diagnostic accuracy for tuberculosis, with its performance influenced by sample type and sequencing platform. These findings support the optimization of tNGS application in specific clinical settings to enhance tuberculosis management.
靶向二代测序（tNGS）是世界卫生组织（WHO）认可的用于结核病诊断的辅助工具。本研究旨在调查tNGS在不同临床场景（人群、样本及结核病类型、平台）中的诊断性能，此前尚未对此进行全面评估。
在PubMed、MEDLINE、Web of Science、万方数据、维普资讯和中国知网进行系统文献检索，以识别2005年1月1日至2024年10月14日期间发表的英文和中文研究，这些研究使用参考菌株以及疑似或确诊结核病患者的原始样本或分离株评估tNGS用于结核病诊断的情况。纳入的研究使用诊断准确性研究质量评估工具-2（QUADAS-2）进行评估，并根据微生物学参考标准和综合参考标准计算诊断准确性。主要结局为合并敏感度和特异度。本研究已在国际前瞻性系统评价注册库（PROSPERO）注册（注册号：CRD42024560190）。
在筛选的3383条记录中，16项研究（共2565名参与者）纳入了荟萃分析。总体而言，tNGS的合并敏感度为0.86（95%CI，0.77-0.91），特异度为0.95（0.86-0.99）。阳性似然比和阴性似然比分别为18.24（5.61-59.26）和0.15（0.09-0.26）。诊断比值比和汇总受试者工作特征曲线下面积分别为120.88（26.93-542.57）和0.95（0.93-0.97）。tNGS的诊断性能因样本类型和测序平台而异，支气管肺泡灌洗液的敏感度最高，为0.91（0.75-0.98），目标纳米孔测序平台的敏感度为0.85（0.80-0.89）。此外，与GeneXpert MTB/RIF相比，tNGS的检测率显著更高（比值比：1.78（1.39-2.29）；p=0.01）。
tNGS对结核病具有较高的诊断准确性，其性能受样本类型和测序平台影响。这些发现支持在特定临床环境中优化tNGS的应用，以加强结核病管理。
