支气管肺泡灌洗液纳米孔测序对肺结核的诊断价值

doi:10.19982/j.issn.1000-6621.20250199

摘要/Abstract

摘要：

目的: 评估支气管肺泡灌洗液(BALF)纳米孔测序对肺结核的诊断价值。方法: 采用前瞻性研究方法,采集2021年10月至2023年8月广西壮族自治区南宁市第四人民医院收治且符合入组标准的577例疑似肺结核患者的支气管肺泡灌洗液样本,分别进行抗酸染色涂片镜检(AFB)、分枝杆菌固体培养、MTB-DNA、GeneXpert MTB/RIF(简称“Xpert”)、纳米孔测序等检测,计算5种检测方法的阳性检出率,并以最终临床诊断为参照标准,对比分析单一方法及联合检测对肺结核的诊断效能。结果: 577例疑似肺结核患者中,473例诊断为肺结核(结核组),104例诊断为非结核疾病(非结核组);经AFB、分枝杆菌固体培养、MTB-DNA、Xpert、纳米孔测序检测的阳性率分别为19.06%(110例)、28.60%(165例)、25.13%(145例)、26.86%(155例)、57.19%(330例);其中,纳米孔测序可独立检出144例肺结核患者。纳米孔测序、分枝杆菌固体培养、Xpert、MTB-DNA、AFB、传统病原学(除纳米孔测序外的其他4种方法联合检测)的检测敏感度分别为68.71%(325/473)、30.44%(144/473)、32.35%(153/473)、30.02%(142/473)、20.08%(95/473)、46.30%(219/473),特异度分别为95.19%(99/104)、79.81%(83/104)、98.08%(102/104)、97.12%(101/104)、85.58%(89/104)、75.96%(79/104),Kappa值分别为0.416、0.048、0.139、0.122、0.024、0.120,AUC值分别为0.820、0.551、0.652、0.636、0.528、0.611。联合诊断策略检测显示,纳米孔测序联合分枝杆菌固体培养、Xpert、MTB-DNA、AFB检测的敏感度分别为72.94%(345/473)、69.98%(331/473)、71.04%(336/473)、70.61%(334/473),特异度分别为76.92%(80/104)、93.27%(97/104)、92.31%(96/104)、82.69%(86/104),Kappa值分别为0.359、0.420、0.427、0.367,AUC值分别为0.751、0.817、0.818、0.768。结论: 相较于其他4种方法,纳米孔测序技术对肺结核的检测效能最佳,且联合MTB-DNA检测的诊断效能也最好,在肺结核诊断中具有显著优势,有望成为肺结核诊断的重要工具。

关键词: 结核,肺, 支气管肺泡灌洗液, 纳米技术, 诊断, 对比研究

Abstract:

Objective: To evaluate the diagnostic value of nanopore sequencing on bronchoalveolar lavage fluid (BALF) for pulmonary tuberculosis(PTB). Methods: A prospective study was conducted. BALF samples were collected from 577 suspected PTB patients who were admitted to the Fourth People’s Hospital of Nanning, Guangxi Zhuang Autonomous Region, from October 2021 to August 2023 and met the inclusion criteria. The samples were subjected to acid-fast bacilli staining smear microscopy (AFB), mycobacterial solid culture, MTB-DNA, GeneXpert MTB/RIF (hereinafter referred to as “Xpert”), and nanopore sequencing. Positive detection rates of the five detection methods were calculated. With the final clinical diagnosis as the reference standard, the diagnostic efficacies of single-method and combined detection method were comparatively analyzed. Results: Among the 577 suspected PTB patients, 473 were diagnosed with PTB, and 104 were diagnosed with non-tuberculous diseases. The positive rates of AFB, mycobacterial solid culture, MTB-DNA, Xpert, and nanopore sequencing were 19.06% (110 cases), 28.60% (165 cases), 25.13% (145 cases), 26.86% (155 cases), and 57.19% (330 cases) respectively. Among them, nanopore sequencing could independently detect 144 positive patients. The sensitivities of nanopore sequencing, mycobacterial solid culture, Xpert, MTB-DNA, AFB, and traditional etiological methods(combined the four methods except nanopore sequencing) were 68.71% (325/473), 30.44% (144/473), 32.35% (153/473), 30.02% (142/473), 20.08% (95/473), and 46.30% (219/473) respectively; the specificities were 95.19% (99/104), 79.81% (83/104), 98.08% (102/104), 97.12% (101/104), 85.58% (89/104), and 75.96% (79/104) respectively; the Kappa values were 0.416, 0.048, 0.139, 0.122, 0.024, and 0.120 respectively; and the AUC values were 0.820, 0.551, 0.652, 0.636, 0.528, and 0.611 respectively. The combined diagnosis strategies showed that sensitivities of nanopore sequencing combined with each one of mycobacterial solid culture, Xpert, MTB-DNA, and AFB were 72.94% (345/473), 69.98% (331/473), 71.04% (336/473), and 70.61% (334/473) respectively; and the specificities were 76.92% (80/104), 93.27% (97/104), 92.31% (96/104), and 82.69% (86/104) respectively; the Kappa values were 0.359, 0.420, 0.427, and 0.367 respectively; and the AUC values were 0.751, 0.817, 0.818, and 0.768 respectively. Conclusion: Compared with the other four methods, nanopore sequencing technology has the best detection efficacy for PTB, and it combined with MTB-DNA test has the best diagnostic efficacy, which shows its significant advantages in diagnosing PTB and is expected to become an important tool for PTB diagnosis.

Key words: Tuberculosis, pulmonary, Bronchoalveolar lavage fluid, Nanotechnology, Diagnosis, Comparative study

中图分类号:

R521
R446.5

朱庆东, 宋畅, 黄爱春, 曾春梅, 李维文, 谭秋清, 赵春艳, 谢周华. 支气管肺泡灌洗液纳米孔测序对肺结核的诊断价值[J]. 中国防痨杂志, 2025, 47(11): 1474-1480. doi: 10.19982/j.issn.1000-6621.20250199

Zhu Qingdong, Song Chang, Huang Aichun, Zeng Chunmei, Li Weiwen, Tan Qiuqing, Zhao Chunyan, Xie Zhouhua. The application value of performing nanopore sequencing on bronchoalveolar lavage fluid in the diagnosis of pulmonary tuberculosis[J]. Chinese Journal of Antituberculosis, 2025, 47(11): 1474-1480. doi: 10.19982/j.issn.1000-6621.20250199

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