Skip to main content

Exploring the microbiota difference of bronchoalveolar lavage fluid between community-acquired pneumonia with or without COPD based on metagenomic sequencing: a retrospective study

Abstract

Background

Community-acquired pneumonia (CAP) patients with chronic obstructive pulmonary disease (COPD) have higher disease severity and mortality compared to those without COPD. However, deep investigation into microbiome distribution of lower respiratory tract of CAP with or without COPD was unknown.

Methods

So we used metagenomic next generation sequencing (mNGS) to explore the microbiome differences between the two groups.

Results

Thirty-six CAP without COPD and 11 CAP with COPD cases were retrieved. Bronchoalveolar lavage fluid (BALF) was collected and analyzed using untargeted mNGS and bioinformatic analysis. mNGS revealed that CAP with COPD group was abundant with Streptococcus, Prevotella, Bordetella at genus level and Cutibacterium acnes, Rothia mucilaginosa, Bordetella genomosp. 6 at species level. While CAP without COPD group was abundant with Ralstonia, Prevotella, Streptococcus at genus level and Ralstonia pickettii, Rothia mucilaginosa, Prevotella melaninogenica at species level. Meanwhile, both alpha and beta microbiome diversity was similar between groups. Linear discriminant analysis found that pa-raburkholderia, corynebacterium tuberculostearicum and staphylococcus hominis were more enriched in CAP without COPD group while the abundance of streptococcus intermedius, streptococcus constellatus, streptococcus milleri, fusarium was higher in CAP with COPD group.

Conclusions

These findings revealed that concomitant COPD have an mild impact on lower airway microbiome of CAP patients.

Peer Review reports

Background

Community acquired pneumonia (CAP) is one of the most frequent infectious causes of death [1,2,3]. Among those CAP patients who were older than 40 years, there are a specific proportion of them concomitant with chronic obstructive pulmonary disease (COPD) [4, 5]. COPD is a progressive pulmonary disorder characterized by persistent airflow limitation and respiratory symptoms, which is now the 3rd leading cause of death [6]. The diagnosis criteria of COPD consists of post-bronchodilator fixed ratio of FEV1/FVC < 70%, which means persistent airflow limitation [7]. A number of well designed large population based studies indicated that COPD is the strongest risk factor for CAP [8,9,10]. Meanwhile, several studies showed that CAP patients got worse disease severity in terms of Pneumonia Severity Index or CURB-65 when concomitant with COPD [11]. Previous studies have attempted to explain the complex etiology of CAP happened in COPD patients, mainly involving lung microbiome, host immunity and pathogen virulence. In general, unlike healthy lung microbiome which harbors low number but a high diversity of bacterium, COPD patients have a dysbiosis lung microbiome, characterized by low diversity [12, 13]. In clinic, considering CAP patients with COPD were at higher risk of deterioration, pulmonary physicians tend to choose a broader-spectrum antibiotics therapy. But the precise lower airway microbiome comparison of CAP with COPD and CAP without COPD is lacking. We would like to know whether there are any differences of the lower airway microbiome distribution between CAP with COPD and CAP without COPD by using metagenomic next generation sequencing (mNGS), an unbiased detection tool of bacteria, eukaryotes and DNA viruses at species level [14, 15].

Methods

Patient selection

Patients whose BALF (bronchoalveolar lavage fluid) had gone through mNGS in the Department of Respiratory Medicine of Shanghai Tenth People’s Hospital (Shanghai, China) between 2019 April to 2021 August were reviewed. Cases which met the diagnosis criteria of community-acquired pneumonia were included, in accordance with the American Thoracic Society/Infectious Diseases Society of America Community-Acquired Pneumonia Guideline [11]. Among those cases, the patients who matched these criteria: a) the presence of a post-bronchodilator FEV1/FVC < 0.7; b)acute deterioration of respiratory symptoms that need additional therapy such as antibiotics) were classified into CAP with COPD group. Otherwise, they would be classified into the CAP without COPD group. Lung carcinoma or immunocompromised patients were excluded.

BALF collection and DNA extraction

Experienced clinicians guided the bronchoscopy to diseased region of the lungs referring to CT results. BALF was typically conducted using serial 20-ml fractions containing 60–100 ml of 0.9% NaCl. 60% of the volume of the lavage was extracted using a gentle syringe suction and transferred to sterile receptacles. Centrifuge the BALF samples at 1000 rpm for 10 min at 4 °C and supernatants and precipitation were separately frozen (− 80 °C) for further analysis. DNA was extracted from the precipitation using a QIAamp® UCP Pathogen DNA Kit (Qiagen) following the manufacturer provided protocol. Human DNA was removed using Benzonase (Qiagen) and Tween20 (Sigma) [16].

Library preparation and sequencing

Ten nanograms DNA samples was used to construct library using Nextera XT DNA Library Prep Kit (Illumina, San Diego, CA) [17]. Library pools were then loaded onto an Illumina Nextseq 550Dx sequencer for 75 cycles of single-end sequencing to generate approximately 20 million reads for each library. Peripheral blood mononuclear cell samples with 105 cells/mL from healthy donors was served as negative controls [17, 18]. DNA-free water also went through DNA extraction and mNGS analysis as a blank control group to assess the degree of background contamination.

Bioinformatics analysis

Low quality reads, adapter contamination, and duplicate reads, reads shorter than 50 bp as well as low complexity reads with default parameters was removed using trimmomatic [19] and Kcomplexity [20]. Human sequence data were identified and excluded referring to a human reference genome (hg38) using Burrows-Wheeler Aligner software [21].

After alignment, multiple indicators were comprehensively evaluated to have the list of suspected microorganisms. Then, microbiota composition profiles were inferred from quality filtered forward reads using Kraken V.2.1.2 and Bracken V.2.6.2 with the k2_pluspf_20210517 database.

Statistical analysis

The site by species counts and relative abundance tables were input into R-base V.4.1.0 for statistical analysis. Alpha diversity of the microbiota profile for each subject was assessed by group at the different level data using the Vegan package in R (version 2.5.7). Different groups were assessed using permutational multi-variate analysis of variance (PERMANOVA) and PCA stat with the Vegan package in R (version 2.5.7). The result of PCOA was stated by the function of dudi.pco in ade4 package in R (version 1.7.18). Associations of specific microbial species or genus with patient parameters were identified using the linear discriminant analysis effect size (LEfSe) [22]. LDA > 2 were considered significantly different.

Results

A total of 36 CAP without COPD and 11 CAP with COPD cases were retrieved (Table 1). In the former group, 24 out of 36 were male while 10 out of 11 in latter group were male gender. As shown in the Table 1, the mean age, CURB 65 and serum biomarkers such as CRP, WBC, NEUT% and the comorbidity rate of hypertension and diabetes mellitus is not significantly different between groups. Body mass index (BMI) was significantly lower in CAP with COPD group (19.96 ± 2.11) compared with CAP without COPD group (23.05 ± 3.1). FEV1/pre was 58.53 ± 16.28 in CAP with COPD and 76.56 ± 20.16 in CAP without COPD. FEV1/FVC was 61.36 ± 9.64 in CAP with COPD and 80.29 ± 3.65 in CAP without COPD.

Table 1 Clinical characteristics of the study population

Table 2 illustrates that the etiology of pneumonia of CAP without COPD group is 38.89% bacteria, 2.78% fungi, 5.55% atypical pathogen and 52.78% unknown, while that for CAP with COPD group is 45.45% bacteria and 54.55% unknown. The conventional cultures result of the BALF is also listed.

Table 2 Etiologic diagnosis of the pneumonia and result of conventional cultures of the bronchoalveolar lavage

Bacterial, viral and fungal composition

Deep metagenome sequencing can totally identified 1012 bacteria, 27 viruses and 18 fungi at genus level, and 3355 bacteria, 71 viruses and 22 fungi at specie level.

Figure 1a showed the top 15 bacteria genus and Fig. 1b showed the top 15 bacteria species respectively in CAP with COPD and CAP without COPD. In CAP with COPD group,the top 10 bacteria genus, from most abundance to least abundance, were Streptococcus, Prevotella, Bordetella, Cutibacterium, Ralstonia, Rothia, Klebsiella, Haemophilus, Veillonella, Staphylococcus. While in CAP group, from most abundance to least abundance, the top 10 bacteria genus were Ralstonia, Prevotella, Streptococcus, Rothia, Staphylococcus, Veillonella, Haemophilus, Cutibacterium, Bordetella etc. In CAP with COPD group, the top 10 bacteria species were Cutibacterium acnes, Rothia mucilaginosa, Bordetella genomosp. 6, etc. While in CAP without COPD group, the top 10 bacteria species were Ralstonia pickettii, Rothia mucilaginosa, Prevotella melaninogenica, etc.

Fig. 1
figure 1

Relative abundance of top 15 microbiome taxa in CAP patients with COPD or without COPD. a Relative abundance of top 15 bacterial genera between CAP patients with COPD or without COPD. b Relative abundance of top 15 bacterial species between CAP patients with COPD or without COPD. c Relative abundance of top 15 fungal and viral genera between CAP patients with COPD or without COPD. d Relative abundance of top 15 fungal and viral species between CAP patients with COPD or without COPD

The most common virus and fungus genus were Gorganvirus and Candida both in CAP with COPD and CAP without COPD group respectively (Fig. 1c). The most common virus and fungus species were Gorganvirus and Candida both in CAP with COPD and CAP without COPD group respectively (Fig. 1d).

Microbiome diversity comparison

Microbiome diversity was compared between CAP with COPD group and CAP without COPD group. Index of shannon, simpson, Richness, ACE, Chao1 were all similar between groups at genus or species level (Fig. 2a-e). Beta diversity was also similar between groups (Fig. 2f-g).

Fig. 2
figure 2

Alpha and beta diversity comparison between CAP patients with COPD or without COPD. (a) Shannon index, (b) Simpson index, (c) Richness index, (d) ACE index and (e) Chao 1 index were similar between CAP patients with COPD or without COPD. PCA (f) and PCoA (g) plot were presented as well

Differential taxa between groups

A LEfSe analysis was applied to figure out specific microbiome whose abundance were statistically different between CAP without COPD and CAP with COPD group. As shown in the Fig. 3, Paraburkholderia, Corynebacterium tuberculostearicum and Staphylococcus hominis were more enriched in CAP without COPD group while the abundance of Streptococcus intermedius, Streptococcus constellatus, Streptococcus milleri, Fusarium was higher in CAP with COPD group.

Fig. 3
figure 3

Linear discriminant analysis revealed differentially abundant bacterial taxa of BALF between CAP patients with COPD or without COPD. a Differences in bacterial genera between CAP patients with COPD or without COPD. b Differences in bacterial species between CAP patients with COPD or without COPD. c Difference in fungal and viral genera between CAP patients with COPD or without COPD

Correlation between the microbiome and clinical indexes

The correlation of microbiome diversity index (Richness, ACE, Chao1, Shannon, Simpson, goods Coverage) and clinical indexes (age, smoking index, BMI, PH, PCO2, PO2, SaO2, white blood cell, neutrophils %, leukomonocyte %, eosinophil, CRP, PCT) was analysed. Figure 4 showed that Simpson index was negatively correlated with age (R = -0.308, p = 0.0355) while ACE was positively correlated with number of eosinophils (R = 0.3, p = 0.0403).

Fig. 4
figure 4

Correlation between BALF microbiome diversity and clinical variables in CAP patients. a Age had a negative correlation with Simpson index(r = -0.308, p = 0.0355). b Number of eosinophils had a positive correlation with ACE index (r = 0.3, p = 0.0403)

Figure 5 revealed potential co-existence and co-exclusion relationship of top 30 bacteria speices and genus separately in each group.

Fig. 5
figure 5

BALF microbiome networks separately in CAP patients with or without COPD. a Heatmap of Spearman correlation of top 30 bacterial genera in CAP patients with COPD. b Heatmap of Spearman correlation of top 30 bacterial species in CAP patients with COPD. c Heatmap of Spearman correlation of top 30 bacterial genera in CAP patients without COPD. d Heatmap of Spearman correlation of top 30 bacterial species in CAP patients without COPD

Discussion

Clinical pneumonologists tend to make a different empirical antibiotic therapy when faced with a CAP patient concomitant with COPD compared a CAP patient without COPD, considering different pathogenic bacteria tendency. However, no one has looked into the microbiome distribution difference of the two group of patients in detail. This is the first article using mNGS techonology to investigate the BALF microbiome distribution of CAP with COPD and CAP without COPD. We found out that 1) there were no statistical difference in microbiome diversity between CAP with COPD and CAP patients without COPD. 2) The specific distribution of microbiome genera and species were different, including the top 10 bacteria taxa, and the abundance of marginal specific microbiome species were different between groups.

The human respiratory system is inhabited by a diverse microbial community. Our study found that the microbial diversity of BALF was similar between CAP patients with COPD and CAP patients without COPD. In our study, we used several different indexes of diversity including shannon, simpson, Richness, ACE, Chao1, each providing different insights into the richness and evenness of microbial communities. The Shannon index measures both richness and evenness within a community. The Simpson index quantifies diversity by considering the probability that two individuals randomly selected from a sample belong to the same species. The ACE index estimates the total number of species present in a community based on abundance data. It takes into account both observed species richness and the number of rare or singleton species that may not have been observed. Chao1 Index predicts the total number of species in a community by considering the number of rare species observed. Different indexes have different calculation method and different emphasis. Qiang Xiao etc. found that compared with healthy individuals, the bacterial alpha diversity in the lower respiratory tract was significantly lower in CAP patients by 16S rDNA gene sequencing of BALF [23]. Loss of diversity has also been observed in COPD, IPF (idiopathic pulmonary fibrosis) and CF (cystic fibrosis), and it has also been described during exacerbations of these diseases [24].

The LDA analysis identified several different bacteria taxa in each group. It is noteworthy that the three bacterial species which had higher abundance in CAP with COPD group: Streptococcus intermedius, Streptococcus constellatus and Streptococcus milleri are subgroups of viridans streptococci. Viridans streptococci are normal inhabitants of mucous membrane-lined cavities of the animals and humans. They are usually found in the upper respiratory tract, all the regions of the gastrointestinal tract, the female genital tract and mostly in the oral cavity. To be more specific, Inhalation of streptococcus intermedius bacteria in the oropharynx may lead to aspiration pneumonia, and may be complicated with lung abscess and empyema. Streptococcus constellatus was associated with abscess formation in the upper body and respiratory tract. It has also been found to be involved with pulmonary exacerbations in cystic fibrosis patients and can lead to toxic shock and limb amputation [25]. As for the coexistence and coexclusion associations observed in the top 30 specific taxa, there is a possibility that needs to be considered that it could happen also in healthy subjects or COPD patients without pneumonia.

Our study explored the lower airway microbiome of CAP with or without COPD deeply by using mNGS. mNGS is an advanced technology in discovering microbiome, including bacteria, eukaryotes and DNA viruses. Meanwhile mNGS allows unbiased detection of microbiome. Furthermore, mNGS is less influenced by antibiotics compared with traditional culture. Due to limited technology, previous microbiome study could only reveal the microbiome at phylum, class or genus level. But mNGS provide us a higher taxonomic resolution at species level [26,27,28]. What’s more, we chose BALF sample to reveal the lower airway microbiome, which is also the closest result to those for the bronchial mucosa [29].

There are some limitations for the study. The study was single centered with small number of enrolled patients. Multi-centered clinical trial is needed in the future. Meanwhile, this is a retrospective study of BALF sample. As the bronchoalveolar liquid is mostly diluted in saline, the effect of dilution can have the impact on the result. Although, all the bronchoscope operator is trained to a standard BALF collection procedure. But there may still be some discrepancies during actual operation.

Conclusions

Although difference of microbiome diversity was no detected between CAP patients with COPD and without COPD. Specific microbiome taxa abundance alteration revealed that concomitant COPD have a mild impact on lower airway microbiome of CAP patients.

Availability of data and materials

Availability of data and materials Raw sequence data have been deposited in the NCBI “Sequence Read Archive” (SRA) under the project accession PRJNA946338. Other data generated or analyzed during this study are included in this article.

Abbreviations

CAP:

Community-acquired pneumonia

COPD:

Chronic obstructive pulmonary dis-ease

mNGS:

Metagenomic next generation sequencing

BALF:

Bronchoalveolar lavage fluid

LEfSe:

Linear discriminant analysis effect size

LDA:

Linear discriminant analysis

BMI:

Body mass index

References

  1. Gadsby NJ, Musher DM. The microbial etiology of community-acquired pneumonia in adults: from classical bacteriology to host transcriptional signatures. Clin Microbiol Rev. 2022;35(4):e0001522.

    Article  PubMed  Google Scholar 

  2. Niederman MS, Torres A. Severe community-acquired pneumonia. Eur Respir Rev. 2022;31(166).

    Article  PubMed  PubMed Central  Google Scholar 

  3. Jiang N, Long QY, Zheng YL, Gao ZC. Advances in epidemiology, etiology, and treatment of community-acquired pneumonia. Zhonghua yu fang yi xue za zhi. 2023;57(1):91–9.

    CAS  PubMed  Google Scholar 

  4. Hsu CW, Suk CW, Hsu YP, Chang JH, Liu CT, Huang SK, Hsu SC. Sphingosine-1-phosphate and CRP as potential combination biomarkers in discrimination of COPD with community-acquired pneumonia and acute exacerbation of COPD. Respir Res. 2022;23(1):63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Yang J, Zhang Q, Zhang J, Ouyang Y, Sun Z, Liu X, Qaio F, Xu LQ, Niu Y, Li J. Exploring the change of host and microorganism in chronic obstructive pulmonary disease patients based on metagenomic and metatranscriptomic sequencing. Front Microbiol. 2022;13:818281.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Stolz D, Mkorombindo T, Schumann DM, Agusti A, Ash SY, Bafadhel M, Bai C, Chalmers JD, Criner GJ, Dharmage SC, et al. Towards the elimination of chronic obstructive pulmonary disease: a Lancet Commission. Lancet (London, England). 2022;400(10356):921–72.

    Article  PubMed  Google Scholar 

  7. Qi C, Sun SW, Xiong XZ. From COPD to lung cancer: mechanisms linking, diagnosis, treatment, and prognosis. Int J Chron Obstruct Pulmon Dis. 2022;17:2603–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Pascual-Guardia S, Amati F, Marin-Corral J, Aliberti S, Gea J, Soni NJ, Rodriguez A, Sibila O, Sanz F, Sotgiu G, et al. Bacterial patterns and empiric antibiotic use in COPD patients with community-acquired pneumonia. Arch Bronconeumol. 2023;59(2):90–100.

    Article  PubMed  Google Scholar 

  9. de Miguel-Diez J, Lopez-Herranz M, Hernandez-Barrera V, de Miguel-Yanes JM, Perez-Farinos N, Wärnberg J, Carabantes-Alarcon D, Jimenez-Garcia R, Lopez-de-Andres A. Community-acquired pneumonia among patients with COPD in Spain from 2016 to 2019 cohort study assessing sex differences in the incidence and outcomes using hospital discharge data. J Clin Med. 2021;10(21):4889.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Yu Y, Liu W, Jiang HL, Mao B. Pneumonia is associated with increased mortality in hospitalized COPD Patients: a systematic review and meta-analysis. Respiration. 2021;100(1):64–76.

    Article  PubMed  Google Scholar 

  11. Metlay JP, Waterer GW, Long AC, Anzueto A, Brozek J, Crothers K, Cooley LA, Dean NC, Fine MJ, Flanders SA, et al. Diagnosis and treatment of adults with community-acquired pneumonia an official clinical practice guideline of the American thoracic society and infectious diseases society of America. Am J Respir Crit Care Med. 2019;200(7):e45–67.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Wang Z, Liu H, Wang F, Yang Y, Wang X, Chen B, Stampfli MR, Zhou H, Shu W, Brightling CE, et al. A refined view of airway microbiome in chronic obstructive pulmonary disease at species and strain-levels. Front Microbiol. 2020;11:1758.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Monsó E. Microbiome in chronic obstructive pulmonary disease. Ann Transl Med. 2017;5(12):251.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Chen Y, Feng W, Ye K, Guo L, Xia H, Guan Y, Chai L, Shi W, Zhai C, Wang J, et al. Application of metagenomic next-generation sequencing in the diagnosis of pulmonary infectious pathogens from bronchoalveolar lavage samples. Front Cell Infect Microbiol. 2021;11:541092.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Pienkowska K, Wiehlmann L, Tümmler B. Airway microbial metagenomics. Microbes Infect. 2018;20(9–10):536–42.

    Article  CAS  PubMed  Google Scholar 

  16. Amar Y, Lagkouvardos I, Silva RL, Ishola OA, Foesel BU, Kublik S, Schöler A, Niedermeier S, Bleuel R, Zink A, et al. Pre-digest of unprotected DNA by Benzonase improves the representation of living skin bacteria and efficiently depletes host DNA. Microbiome. 2021;9(1):123.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Miller S, Naccache SN, Samayoa E, Messacar K, Arevalo S, Federman S, Stryke D, Pham E, Fung B, Bolosky WJ, et al. Laboratory validation of a clinical metagenomic sequencing assay for pathogen detection in cerebrospinal fluid. Genome Res. 2019;29(5):831–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Li H, Gao H, Meng H, Wang Q, Li S, Chen H, Li Y, Wang H. Detection of pulmonary infectious pathogens from lung biopsy tissues by metagenomic next-generation sequencing. Front Cell Infect Microbiol. 2018;8:205.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Xu Y, Kang L, Shen Z, Li X, Wu W, Ma W, Fang C, Yang F, Jiang X, Gong S, et al. Dynamics of severe acute respiratory syndrome coronavirus 2 genome variants in the feces during convalescence. J Genet Genomics. 2020;47(10):610–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics (Oxford, England). 2014;30(15):2114–20.

    CAS  PubMed  Google Scholar 

  21. Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics (Oxford, England). 2009;25(14):1754–60.

    CAS  PubMed  Google Scholar 

  22. Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C. Metagenomic biomarker discovery and explanation. Genome Biol. 2011;12(6):R60.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Xiao Q, Tan S, Liu C, Liu B, Li Y, Guo Y, Hu P, Su Z, Chen S, Lei W, et al. Characterization of the microbiome and host’s metabolites of the lower respiratory tract during acute community-acquired pneumonia identifies potential novel markers. Infect Drug Resist. 2023;16:581–94.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Faner R, Sibila O, Agustí A, Bernasconi E, Chalmers JD, Huffnagle GB, Manichanh C, Molyneaux PL, Paredes R, Brocal VP, et al. The microbiome in respiratory medicine: current challenges and future perspectives. Eur Respir J. 2017;49(4):1602086.

    Article  PubMed  Google Scholar 

  25. Sibley CD, Parkins MD, Rabin HR, Duan K, Norgaard JC, Surette MG. A polymicrobial perspective of pulmonary infections exposes an enigmatic pathogen in cystic fibrosis patients. Proc Natl Acad Sci U S A. 2008;105(39):15070–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Gu W, Miller S, Chiu CY. Clinical metagenomic next-generation sequencing for pathogen detection. Annu Rev Pathol. 2019;14:319–38.

    Article  CAS  PubMed  Google Scholar 

  27. Wensel CR, Pluznick JL, Salzberg SL, Sears CL. Next-generation sequencing: insights to advance clinical investigations of the microbiome. J Clin Invest. 2022;132(7):e154944.

  28. de Vries JJC, Brown JR, Couto N, Beer M, Le Mercier P, Sidorov I, Papa A, Fischer N, Oude Munnink BB, Rodriquez C, et al. Recommendations for the introduction of metagenomic next-generation sequencing in clinical virology, part II: bioinformatic analysis and reporting. J Clin Virol. 2021;138:104812.

    Article  PubMed  Google Scholar 

  29. Cabrera-Rubio R, Garcia-Núñez M, Setó L, Antó JM, Moya A, Monsó E, Mira A. Microbiome diversity in the bronchial tracts of patients with chronic obstructive pulmonary disease. J Clin Microbiol. 2012;50(11):3562–8.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

None.

Funding

This work was supported by Shanghai Science and Technology (18410720900) and Fundamental Research Funds for the Central Universities(22120240288). The funder has no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Authors and Affiliations

Authors

Contributions

Conception and design, CHW, MT, SSX; Sample acquisition, WL, QHX, LFJ ; Sample processing, BBW; Data analysis, MT. Drafting of manuscript, BBW; Data interpretation, manuscript revision, final approval, all. All authors read and approved the final manuscript

Corresponding authors

Correspondence to Shuanshuan Xie or Changhui Wang.

Ethics declarations

Ethics approval and consent to participate

The institutional review board for human studies at Shanghai Tenth People’s Hospital approved the protocol and written informed consent was obtained from the subjects. All methods were carried out in accordance with the Declaration of Helsinki.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, B., Tan, M., Li, W. et al. Exploring the microbiota difference of bronchoalveolar lavage fluid between community-acquired pneumonia with or without COPD based on metagenomic sequencing: a retrospective study. BMC Pulm Med 24, 278 (2024). https://doi.org/10.1186/s12890-024-03087-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12890-024-03087-6

Keywords