循环游离DNA检测在结核病诊断中的研究进展

doi:10.19871/j.cnki.xfcrbzz.2026.04.015

摘要/Abstract

摘要： 结核病是全球重大公共卫生挑战,现有诊断方法在敏感性、检测速度和生物安全性等方面仍然存在不足。因此,亟需开发更加精准、高效、快捷的结核病诊断新策略。循环游离DNA（circulating cell-free DNA,cfDNA）是一种存在于人体血液循环、游离于细胞外的高度片段化DNA,已成为极具潜力的新型生物标志物。结核分枝杆菌cfDNA（mycobacterium tuberculosis cfDNA,MTB-cfDNA）可证明宿主体内存在病原体,在结核病的早期与精准诊断方面显示潜力。本综述围绕MTB-cfDNA在结核病诊断中的研究进展,从技术比较、临床诊断应用、样本类型和技术优化方面,系统梳理了MTB-cfDNA检测技术在结核病诊断中的应用进展,并探讨不同类型临床样本中MTB-cfDNA的处理技术、检出效率与适用场景,旨在为结核病的快速精准诊断提供有益支持。

关键词: 结核病, 循环游离DNA, 结核分枝杆菌循环游离DNA, 循环游离DNA诊断技术

Abstract: Tuberculosis remains a major global public health challenge. Although molecular detection techniques have advanced in recent years, existing diagnostic methods still have limitations in terms of sensitivity, detection speed, and biosafety. Therefore, there is an urgent need to develop more precise, efficient, and rapid new diagnostic strategies for tuberculosis. Circulating cell-free DNA (cfDNA), a highly fragmented DNA present in human blood circulation and free from cells, has emerged as a promising novel biomarker. Among these, mycobacterium tuberculosis cfDNA (MTB-cfDNA) can indicate the presence of the pathogen in the host and shows potential for the early and precise diagnosis of tuberculosis. This review systematically summarizes the application progress of MTB-cfDNA detection technology in tuberculosis diagnosis from the perspectives of technical comparison, clinical diagnostic application, sample types, and technological optimization. It further discusses the processing techniques, detection efficiency, and applicable scenarios of MTB-cfDNA in different clinical sample types, aiming to provide a theoretical basis for the rapid and accurate diagnosis of tuberculosis..

Key words: Tuberculosis, Circulating cell-free DNA, Mycobacterium tuberculosis circulating cell-free DNA, Circulating cell-free DNA diagnostic technology

中图分类号:

R521
R378.91

张羽, 黄华, 王卫华, 谭英. 循环游离DNA检测在结核病诊断中的研究进展[J]. 新发传染病电子杂志, 2026, 11(4): 95-100.

Zhang Yu, Huang Hua, Wang Weihua, Tan Ying. Research progress in circulating cell-free DNA detection for tuberculosis diagnosis[J]. Electronic Journal of Emerging Infectious Diseases, 2026, 11(4): 95-100.

参考文献

[1] 何华伟, 黄爱春, 李纬, 等. WHO 2025年全球结核病报告:全球与中国关键数据分析[J/CD]. 新发传染病电子杂志, 2025, 10(6): 1-9.
[2] Matteo M J, Latini M C,Martinovic D N, et al.Update of diagnostic methods in tuberculosis (TB)[J]. Rev Argent Microbiol, 2025, 57(1): 49-53.
[3] Dong B, He Z, Li Y, et al.Improved Conventional and New Approaches in the Diagnosis of Tuberculosis[J]. Front Microbiol, 2022, 13: 924410.
[4] 中国医疗保健国际交流促进会临床微生物与感染分会, 中华医学会检验医学分会临床微生物学组. 综合医院结核分枝杆菌感染实验室检查共识[J].中华检验医学杂志, 2022, 45(4): 343-353.
[5] Heitzer E, Auinger L, Speicher M R.Cell-Free DNA and Apoptosis: How Dead Cells Inform About the Living[J]. Trends Mol Med, 2020, 26(5): 519-528.
[6] Mandel P, Metais P.Les acides nucléiques du plasma sanguin chez l'homme [Nuclear Acids In Human Blood Plasma[J]. C R Seances Soc Biol Fil, 1948, 142(3-4): 241-243.
[7] Hu Z, Chen H, Long Y, et al.The main sources of circulating cell-free DNA: Apoptosis, necrosis and active secretion[J]. Crit Rev Oncol Hematol, 2021, 157: 103166.
[8] Fernandez-Carballo BL, Broger T, Wyss R, et al.Toward the Development of a Circulating Free DNA-Based In Vitro Diagnostic Test for Infectious Diseases: a Review of Evidence for Tuberculosis[J]. J Clin Microbiol, 2019, 57(4): e01234-e01218.
[9] 高聪聪, 曾思恩. 数字PCR技术在体液中检测恶性肿瘤的研究进展[J].华夏医学, 2023, 36(1) : 171-177.
[10] YANG J, Han X, Liu A, et al.Use of Digital Droplet PCR to Detect Mycobacterium tuberculosis DNA in Whole Blood-Derived DNA Samples from Patients with Pulmonary and Extrapulmonary Tuberculosis[J]. Front Cell Infect Microbiol, 2017, 7: 369.
[11] Satam H, Joshi K, Mangrolia U, et al.Next-Generation Sequencing Technology: Current Trends and Advancements[J]. Biology (Basel), 2024, 13(5): 286.
[12] Zein-Eddine R, Refregier G, Cervantes J, et al.The future of CRISPR in Mycobacterium tuberculosis infection[J]. J Biomed Sci, 2023, 30(1): 34.
[13] Zhang S, Xu Y, Huang M, et al.Enhanced diagnosis of tuberculous pleurisy using a multiplex droplet digital PCR assay targeting circulating mycobacterial DNA[J]. Thorax, 2025, 223553.
[14] Xu F, Du W, Li C, et al.Evaluation of droplet digital polymerase chain reaction by detecting cell-free deoxyribonucleic acid in pleural effusion for the diagnosis of tuberculous pleurisy: a multicentre cohort study[J]. Clin Microbiol Infect, 2024, 30(9): 1164-1169.
[15] Quan S, Tian X, Sun Y, et al.Cell-free DNA next-generation sequencing for Mycobacterium tuberculosis obtained from plasma of children with active tuberculosis[J]. BMC Pediatr, 2025, 25(1): 164.
[16] Oreskovic A, Waalkes A, Holmes E A, et al.Characterizing the molecular composition and diagnostic potential of Mycobacterium tuberculosis urinary cell-free DNA using next-generation sequencing[J]. Int J Infect Dis, 2021, 112: 330-337.
[17] Sharma K, Sharma M, Shree R, et al.Diagnosing tuberculous meningitis from cell-free DNA by multi-targeted real-time PCR: An experience from 170 cases[J]. Tuberculosis (Edinb), 2025, 155: 102705.
[18] Sharma K, Sharma M, Joshi H, et al.Evaluation of Mycobacterium Tuberculosis Derived Cell-Free DNA-Based Multi-Targeted Real-Time PCR from Vitreous Fluid (VF) Samples to Diagnose Ocular Tuberculosis[J]. Ocul Immunol Inflamm, 2025, 33(9): 1907-1912.
[19] Ayalew S, Wegayehu T, Wondale B, et al.Multigene Mycobacterium tuberculosis cell-free DNA assay[J]. Int J Tuberc Lung Dis, 2025, 29(1): 13-19.
[20] Huang Z, Lacuures S M, Kay A W, et al.CRISPR detection of circulating cell-free Mycobacterium tuberculosis DNA in adults and children, including children with HIV: a molecular diagnostics study[J]. Lancet Microbe, 2022, 3(7): e482-e492.
[21] Du W L, Liang J Q, Yang X T, et al.Accuracy of cell-free Mycobacterium tuberculosis DNA testing in pleural effusion for diagnosing tuberculous pleurisy: a multicenter cross-sectional study[J]. Mil Med Res, 2024, 11(1): 60.
[22] Yu G, Shen Y, Ye B, et al.Diagnostic accuracy of Mycobacterium tuberculosis cell-free DNA for tuberculosis: A systematic review and meta-analysis[J]. PLoS One, 2021, 16(6): e0253658.
[23] Burnham P, Dadhania D, Heyang M, et al.Urinary cell-free DNA is a versatile analyte for monitoring infections of the urinary tract[J]. Nat Commun, 2018, 9(1): 2412.
[24] Lo YYD, Han DSC, Jiang P, et al. Epigenetics, fragmentomics,topology of cell-free DNA in liquid biopsies[J]. Science, 2021, 372(6538): eaaw3616.
[25] Chang A, Loy C J, Eweis-Labolle D, et al.Circulating cell-free RNA in blood as a host response biomarker for detection of tuberculosis[J]. Nat Commun, 2024, 15(1): 4949.
[26] Gahlawat A W, Lenhardt J, Witte T, et al.Evaluation of Storage Tubes for Combined Analysis of Circulating Nucleic Acids in Liquid Biopsies[J]. Int J Mol Sci, 2019, 20(3): 704.
[27] Ushio R, Yamamoto M, Nakashima K, et al.Digital PCR assay detection of circulating Mycobacterium tuberculosis DNA in pulmonary tuberculosis patient plasma[J]. Tuberculosis (Edinb), 2016, 99: 47-53.
[28] Park J H, Koo B, Kim M J, et al.Utility of plasma cell-free DNA detection using homobifunctional imidoesters using a microfluidic system for diagnosing active tuberculosis[J]. Infect Dis (Lond), 2022, 54(1): 46-52.
[29] Thakku S G, Lirette J, Murugesan K, et al.Genome-wide tiled detection of circulating Mycobacterium tuberculosis cell-free DNA using Cas13[J]. Nat Commun, 2023, 14(1): 1803.
[30] Li G, Cannon K, Sisniega C, et al.Cell-free DNA blood test for the diagnosis of pediatric tuberculous meningitis[J]. J Clin Tuberc Other Mycobact Dis, 2024, 35: 100421.
[31] Youngquist B M, Saliba J, Kim Y, et al. Rapid tuberculosis diagnosis from respiratory or blood samples by a low cost, portable lab-in-tube assay[J]. Sci Transl Med, 2025, 17(793): eadp6411.
[32] Meng Y, Qu R, Wang Y, et al.Target-activated DNAzyme and ion-functionalized DNA nanospheres enable dual cfDNA homogeneous electrochemical analysis for tuberculosis diagnosis[J]. Chem Eng J, 2025, 520: 166407.
[33] Oreskovic A, Panpradist N, Marangu D, et al.Diagnosing Pulmonary Tuberculosis by Using Sequence-Specific Purification of Urine Cell-Free DNA[J]. J Clin Microbiol, 2021, 59(8): e0007421.
[34] Sharma P, Anthwal D, Kumari P, et al.Utility of circulating cell-free Mycobacterium tuberculosis DNA for the improved diagnosis of abdominal tuberculosis[J]. PLoS One, 2020, 15(8): e0238119.
[35] Labugger I, Heyckendorf J, Dees S, et al.Detection of transrenal DNA for the diagnosis of pulmonary tuberculosis and treatment monitoring[J]. Infection, 2017, 45(3): 269-276.
[36] Patel K, Nagel M, Wesolowski M, et al.Evaluation of a Urine-Based Rapid Molecular Diagnostic Test with Potential to Be Used at Point-of-Care for Pulmonary Tuberculosis: Cape Town Cohort[J]. J Mol Diagn, 2018, 20(2): 215-224.
[37] Tschan Y, Sasamalo M, Hiza H, et al.Diagnostic accuracy of a sequence-specific Mtb-DNA hybridization assay in urine: a case-control study including subclinical TB cases[J]. Microbiol Spectr, 2024, 12(6): e0042624.
[38] Wu H, Ji H, Yang W, et al.Liquid biopsy using ascitic fluid and pleural effusion supernatants for genomic profiling in gastrointestinal and lung cancers[J]. BMC Cancer, 2022, 22(1): 1020.
[39] Villatoro S, Mayo-De-Las-Casas C, Jordana-Ariza N, et al. Prospective detection of mutations in cerebrospinal fluid, pleural effusion, and ascites of advanced cancer patients to guide treatment decisions[J]. Mol Oncol, 2019, 13(12): 2633-2645.
[40] Iser F, Hinz F, Hoffmann D C, et al.Cerebrospinal Fluid cfDNA Sequencing for Classification of Central Nervous System Glioma[J]. Clin Cancer Res, 2024, 30(14): 2974-2985.
[41] 寿娟,谢青梅,易伟,等.胸水游离DNA的结核杆菌检测在结核病诊断中的价值[J].中华病理学杂志,2018, 47(6): 465-467.
[42] Yang X, Che N, Duan H, et al. Cell-free Mycobacterium tuberculosis DNA test in pleural effusion for tuberculous pleurisy: a diagnostic accuracy study[J]. Clin Microbiol Infect, 2020, 26(8): 1089.e1-1089.e6.
[43] Shao L, Qiu C, Zheng L, et al.Comparison of diagnostic accuracy of the GeneXpert Ultra and cell-free nucleic acid assay for tuberculous meningitis: A multicentre prospective study[J]. Int J Infect Dis, 2020, 98: 441-446.
[44] Chen S, Wang C, Zou Y, et al.Tuberculosis-targeted next-generation sequencing and machine learning: An ultrasensitive diagnostic strategy for paucibacillary pulmonary tuberculosis and tuberculous meningitis[J]. Clin Chim Acta, 2024, 553: 117697.