基于宏基因组学对肺结核合并下呼吸道感染患者支气管肺泡灌洗液微生物谱特征分析

doi:10.19871/j.cnki.xfcrbzz.2026.04.001

摘要/Abstract

摘要： 目的探讨肺结核合并下呼吸道感染（pulmonary tuberculosis complicated by lower respiratory tract infection,PTB-LRTI）患者支气管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）微生物谱特征,初步阐释菌群在疾病中的潜在参与机制,为PTB-LRTI的诊断提供理论依据。方法收集2023年6月至2024年12月甘肃省人民医院诊治的98例患者BALF样本,其中PTB-LRTI组60例,LRTI组38例。采用宏基因组测序技术分析BALF微生物组成,通过α多样性分析、线性判别分析效应量（linear discriminant analysis effect size,LEfSe）差异分析及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）功能注释比较两组微生物群落结构及功能差异。结果与LRTI组相比,PTB-LRTI组BALF微生物谱α多样性显著升高（P<0.05）。PTB-LRTI组厚壁菌门、放线菌门及拟杆菌门丰度增加,分枝杆菌属、诺卡菌属及结核分枝杆菌、皮疽诺卡菌显著富集;LRTI组主要富集变形菌门及链球菌属、克雷伯菌属和嗜血杆菌属。PTB-LRTI组鉴定物种数高于LRTI组（396种对145种）。LEfSe分析显示两组在多个分类水平存在显著差异。KEGG功能分析提示萜类主干生物合成、环腺苷酸信号通路及磷酸生物合成等功能存在差异（P<0.05）。结论 PTB-LRTI患者BALF微生物群落结构及功能谱呈现显著改变,以厚壁菌门、放线菌门及分枝杆菌属富集为特征。差异菌群与萜类主干生物合成等代谢功能紊乱相关,提示BALF微生物谱分析可为PTB-LRTI的病原学诊断及机制研究提供新的理论依据。

关键词: 肺结核, 下呼吸道感染, 宏基因组二代测序, 支气管肺泡灌洗液, 微生物谱

Abstract: Objective To investigate the microbial profiling characteristics of bronchoalveolar lavage fluid (BALF) in patients with pulmonary tuberculosis complicated by lower respiratory tract infection (PTB-LRTI), and to preliminarily elucidate the potential mechanisms of microbiota involvement in the disease, thereby providing a theoretical basis for the diagnosis of PTB-LRTI. Method BALF samples were collected from 98 patients treated at Gansu Provincial People's Hospital from June 2023 to December 2024, including 60 patients in the PTB-LRTI group and 38 patients in the LRTI group. Metagenomic next-generation sequencing (mNGS) was employed to analyze the microbial composition of BALF. The microbial community structure and functional differences between the two groups were compared using α-diversity analysis, linear discriminant analysis effect size (LEfSe) analysis, and Kyoto encyclopedia of genes and genomes CCCCCC(KEGG) functional annotation. Result Compared with the LRTI group, the PTB-LRTI group showed significantly higher α-diversity of the BALF microbial profile (P<0.05). The abundances of Firmicutes, Actinobacteria, and Bacteroidetes were increased in the PTB-LRTI group, with significant enrichment of Mycobacterium, Nocardia, Mycobacterium tuberculosis, and Nocardia farcinica. In the LRTI group, Proteobacteria, as well as Streptococcus, Klebsiella, and Haemophilus, were predominantly enriched. The number of species identified in the PTB-LRTI group was higher than that in the LRTI group (396 vs. 145 species). LEfSe analysis revealed significant differences between the two groups at multiple taxonomic levels. KEGG functional analysis showed significant differences in terpenoid backbone biosynthesis, cyclic adenosine monophosphate signaling pathway, and phosphonate biosynthesis between the two groups (P<0.05). Conclusion The BALF microbial community structure and functional profiles in patients with PTB-LRTI are significantly altered, characterized by enrichment of Firmicutes, Actinobacteria, and Mycobacterium. The differential microbiota are associated with disturbances in metabolic functions, including terpenoid backbone biosynthesis, suggesting that BALF microbial profiling analysis may provide a novel theoretical basis for the etiological diagnosis and mechanistic research of PTB-LRTI.

Key words: Tuberculosis, Lower respiratory tract infection, Metagenomic next-generation sequencing, Bronchoalveolar lavage fluid, Microbial profiles

中图分类号:

R446

刘钰, 唐君霞, 赵千千, 刘宗霖, 贾丽娜, 田悦由, 虎维东. 基于宏基因组学对肺结核合并下呼吸道感染患者支气管肺泡灌洗液微生物谱特征分析[J]. 新发传染病电子杂志, 2026, 11(4): 1-12.

Liu Yu, Tang Junxia, Zhao Qianqian, Liu Zonglin, Jia Lina, Tian Yueyou, Hu Weidong. Analysis of the microbial profile of bronchoalveolar lavage fluid in patients with pulmonary tuberculosis complicated by lower respiratory tract infection using metagenomic sequencing[J]. Electronic Journal of Emerging Infectious Diseases, 2026, 11(4): 1-12.

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] Swaminathan N, Perloff S R, Zuckerman J M.Prevention of Mycobacterium tuberculosis Transmission in Health Care Settings[J]. Infect Dis Clin North Am, 2021, 35(4): 1013-1025.
[3] Cavallazzi R, Ramirez J A.How and when to manage respiratory infections out of hospital[J]. Eur Respir Rev, 2022, 31(166): 220092.
[4] Murray C J L. The Global Burden of Disease Study at 30 years[J].Nat Med, 2022, 28(10): 2019-2026.
[5] Ruan Z, Qi J, Qian Z M, et al.Disease burden and attributable risk factors of respiratory infections in China from 1990 to 2019[J].Lancet Reg Health West Pac, 2021, 11: 100153.
[6] Saladié M, Caparrós-Martín J A, Agudelo-Romero P, et al. Microbiomic Analysis on Low Abundant Respiratory Biomass Samples; Improved Recovery of Microbial DNA From Bronchoalveolar Lavage Fluid[J]. Front Microbiol, 2020, 11: 572504.
[7] Chiu C Y, Miller S A.Clinical metagenomics[J]. Nat Rev Genet, 2019, 20(6): 341-355.
[8] Piombo E, Abdelfattah A, Droby S, et al.Metagenomics Approaches for the Detection and Surveillance of Emerging and Recurrent Plant Pathogens[J]. Microorganisms, 2021, 9(1): 188.
[9] 中华人民共和国国家卫生和计划生育委员会. 肺结核诊断 WS 288—2017[J]. 中国感染控制杂志, 2018, 17(7):642-652.
[10] 中华医学会呼吸病学分会.中国成人社区获得性肺炎诊断和治疗指南(2016年版)[J]. 中华结核和呼吸杂志, 2016, 39(4): 253-279.
[11] 周道银,吴茅,许绍强,等.支气管肺泡灌洗液细胞形态学检验中国专家共识(2020)[J]. 现代检验医学杂志, 2020, 35(6): 4-8.
[12] Delgado L F, Andersson A F.Evaluating metagenomic assembly approaches for biome-specific gene catalogues[J]. Microbiome, 2022, 10(1): 72.
[13] 邹鑫,彭梅芳,阚新锐,等.双叉犀金龟(Trypoxylus dichotomus)幼虫肠道微生物多样性及纤维素降解菌筛选[J]. 微生物学报, 2025, 65(1): 225-238.
[14] Konechnyi Y, Rumynska T, Yushyn I, et al.A New 4-Thiazolidinone Derivative (Les-6490) as a Gut Microbiota Modulator: Antimicrobial and Prebiotic Perspectives[J]. Antibiotics (Basel), 2024, 13(4): 291.
[15] Cai X, Luo Y, Zhang Y, et al.Airway microecology in rifampicin-resistant and rifampicin-sensitive pulmonary tuberculosis patients[J]. BMC Microbiol, 2022, 22(1): 286.
[16] Liu Y, Zhang J, Leng G, et al.Mycobacterium tuberculosis Rv1987 protein attenuates inflammatory response and consequently alters microbiota in mouse lung[J]. Front Cell Infect Microbiol, 2023, 13: 1256866.
[17] Greve D, Moter A, Kleinschmidt M C, et al.Rothia aeria and Rothia dentocariosa as biofilm builders in infective endocarditis[J]. Int J Med Microbiol, 2021, 311(2): 151478.
[18] Gonzalez L M, Venkatesan R, Amador P, et al.Rangaswamy B. TB or Not TB: Lung Nocardiosis, a Tuberculosis Mimicker[J]. Cureus, 2024, 16(3): e55412.
[19] Vos LM, Bruyndonckx R, Zuithoff NPA, et al.GRACE Consortium. Lower respiratory tract infection in the community: associations between viral aetiology and illness course[J]. Clin Microbiol Infect, 2021, 27(1): 96-104.
[20] Aliberti S, Dela Cruz C S, Amati F, et al. Community-acquired pneumonia[J]. Lancet, 2021, 398(10303): 906-919.
[21] Xiao G, Cai Z, Guo Q, et al.Insights into the Unique Lung Microbiota Profile of Pulmonary Tuberculosis Patients Using Metagenomic Next-Generation Sequencing[J]. Microbiol Spectr, 2022, 10(1): e0190121.
[22] Vázquez-Pérez J A, Carrillo C O, Iñiguez-García M A, et al. Alveolar microbiota profile in patients with human pulmonary tuberculosis and interstitial pneumonia[J]. Microb Pathog, 2020, 139: 103851.
[23] Ferluga J, Yasmin H, Al-Ahdal M N, et al. Natural and trained innate immunity against Mycobacterium tuberculosis[J]. Immunobiology, 2020, 225(3): 151951.
[24] Chen Y, Hu W.Co-infection with Mycobacterium tuberculosis and Nocardia farcinica in a COPD patient: a case report[J]. BMC Pulm Med, 2023, 23(1): 136.
[25] Qin L, Xu J, Chen J, et al. Cell-autonomous targeting of arabinogalactan by host immune factors inhibits mycobacterial growth[J]. Elife, 2024, 13: RP92737.
[26] Boronat A, Rodríguez-Concepción M.Terpenoid biosynthesis in prokaryotes[J]. Adv Biochem Eng Biotechnol, 2015, 148:3-18.
[27] Niekamp P, Guzman G, Leier H C, et al.Sphingomyelin Biosynthesis Is Essential for Phagocytic Signaling during Mycobacterium tuberculosis Host Cell Entry[J]. mBio, 2021, 12(1): e03141-20.
[28] Cheng X, Ning J, Xu X, et al.The role of bacterial cyclic di-adenosine monophosphate in the host immune response[J]. Front Microbiol, 2022, 13: 958133.
[29] Liu N, Pang X, Zhang H, et al.The cGAS-STING Pathway in Bacterial Infection and Bacterial Immunity[J]. Front Immunol, 2022, 12:814709.