Diversity of nontuberculous mycobacterial infections in Kuwait: rapid identification and differentiation of Mycobacterium species by multiplex PCR, INNO-LiPA Mycobacteria v2 assay and PCR-sequencing of rDNA
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PROVIDER: PRJEB29664 | ENA |
REPOSITORIES: ENA
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Project description:ObjectiveNontuberculous mycobacteria (NTM) often cause disease that is clinically indistinguishable from tuberculosis. Specific identification is important as treatment varies according to Mycobacterium species causing the infection. This study used multiplex PCR (mPCR) assay for rapid differentiation of mycobacterial growth indicator tube 960 system (MGIT) cultures as Mycobacterium tuberculosis (MTB) or NTM together with INNO LiPA Mycobacteria v2 assay (LiPA) and/or PCR sequencing of rDNA for species-specific identification of selected MTB and all NTM isolates in Kuwait.Materials and methodsDNA was extracted from MGIT cultures (n = 1,033) grown from 664 pulmonary and 369 extrapulmonary specimens from 1,033 suspected tuberculosis patients. mPCR was performed to differentiate MTB from NTM. LiPA was performed and results were interpreted according to kit instructions. rDNA was amplified and sequenced by using panmycobacterial primers.ResultsmPCR identified 979 isolates as MTB, 53 as NTM and 1 isolate as mixed culture. LiPA and/or PCR sequencing confirmed 112 of 979 selected isolates as MTB. Mixed culture contained M. tuberculosis and M. fortuitum. LiPA yielded 12 patterns and identified 10 species/species complexes among 47 NTM, M. kansasii + M. scrofulaceum in one culture and 5 isolates only at genus level. PCR sequencing yielded more specific identification for 22 isolates at the species/subspecies level.ConclusionsmPCR rapidly differentiated MTB from NTM. LiPA identified 44 of 52 NTM isolates at the species/species complex level and 2 mixed cultures. PCR sequencing yielded more specific identification at the species/subspecies level. Rapid differentiation as MTB or NTM by mPCR, followed by species-specific NTM identification by LiPA/PCR sequencing is suitable for the proper management of mycobacterial infections in Kuwait.
| S-EPMC6597937 | biostudies-literature
Project description:Nontuberculous mycobacteria Metagenome
| PRJNA779755 | ENA
Project description:The incidence of pulmonary nontuberculous mycobacterial (PNTM) disease is increasing, but host susceptibility factors are not fully understood. We infected air-liquid interface (ALI) primary respiratory epithelial cell cultures with Mycobacterium avium complex (MAC) or Mycobacterium abscessus (MAB) and performed transcriptome sequencing (RNA-Seq) to identify relevant gene expression differences. We used cells from 4 different donors in order to try to obtain generalizable data. The differentiated respiratory epithelial cells in ALI were infected with MAC or MAB at MOI of 100:1 or 1000:1, and RNA-seq was performed at 1 and 3 days after infection. We found downregulation of ciliary genes, including several identified with polymorphisms in previous PNTM cohorts. The cytokine IL-32, the superpathway of cholesterol biosynthesis and downstream targets within the IL-17 signaling pathway were all elevated. The integrin signaling pathway was more upregulated by MAB than MAC infection. Working with primary respiratory epithelial cells infected with nontuberculous mycobacteria at ALI, we identified ciliary function, cholesterol biosynthesis, chemokine production and the IL-17 pathway as major targets of host responses to infection. Some of these pathways may be amenable to therapeutic manipulation.
2017-10-13 | GSE93526 | GEO
Project description:We performed Spatial transcriptomics (Visium) using the 10X Genomics platform to investigate the role of the autophagy-related protein ATG7 in the context of nontuberculous mycobacterial (NTM) infection. ATG7 in innate immune cells plays a critical role in controlling NTM infection and protecting lung tissue from pathological inflammation. This study represents single-cell analysis of Mice deletion of Atg7 in innate immune cells and reveals the importance of ATG7 in mediating antimicrobial responses during infection and its essential role in host defense mechanisms against NTM.
2025-07-09 | GSE297621 | GEO
Project description:We performed single-cell RNA sequencing (scRNA-seq) using the 10X Genomics platform to investigate the role of the autophagy-related protein ATG7 in the context of nontuberculous mycobacterial (NTM) infection. ATG7 in innate immune cells plays a critical role in controlling NTM infection and protecting lung tissue from pathological inflammation. This study represents single-cell analysis of Mice deletion of Atg7 in innate immune cells and reveals the importance of ATG7 in mediating antimicrobial responses during infection and its essential role in host defense mechanisms against NTM.
2025-07-09 | GSE297620 | GEO
Project description:Genome Sequences and Assembly of Clinical and Environmental Isolates of Nontuberculous Mycobacteria
| PRJNA944390 | ENA
Project description:Profiling Mycobacterial Communities in Pulmonary Nontuberculous Mycobacterial Disease
| PRJEB22153 | ENA
Project description:Nontuberculous mycobacterial pulmonary diseases (NTM-PD) are becoming increasingly prevalent and incident, and heightened antibiotic resistance presents a considerable challenge to clinicians. Despite this, our understanding of the factor(s) contributing to host defense against NTM-PD remains incomplete. Herein, we report that the myeloid autophagy-related gene (ATG) 7 promotes host protective responses against NTM-PD by mitigating excessive pathological inflammation, oxidative stress, and cell death associated with neutrophil infiltration. Patients with NTM-PD displayed a notable reduction in ATG7 expression in peripheral blood mononuclear cells and in necrotic lesions at disease sites. Mice with deletion of Atg7 specifically in myeloid cells (Atg7 cKO) exhibited a significant increase in lung bacterial load during infection. Spatial and bulk RNA-seq analyses, as well as biological experiments, revealed heightened myeloid cell infiltration, excessive inflammation, and mitochondrial damage in Atg7 cKO lung tissues. Furthermore, ｍyeloid Atg7-deficient mice showed upregulated apoptosis, necrosis, and GSDME-associated cell death, along with the formation of neutrophil extracellular traps, in the lung tissues during NTM-PD. However, Atg7-deficient macrophages showed only marginal differences in inflammation and cell death in vitro during NTM infection. Our findings demonstrate that myeloid ATG7 plays a major role in non-cell-autonomous protection against NTM-PD by alleviating neutrophil-associated pathological inflammation and cell death.
2025-05-20 | GSE267176 | GEO
Project description:The immunologic features of nontuberculous mycobacterial pulmonary disease (NTM-PD) are largely unclear. This study investigated the immunologic features of NTM-PD using digital spatial profiling techniques. Lung tissues obtained from six patients with NTM-PD between January 1, 2006, and December 31, 2020, at Seoul National University Hospital were subjected to RNA sequencing. Cores from the peribronchial and fibrotic stromal areas were stained with CD3, CD68, and DNASyto13, and gene expression at the whole-transcriptome level was quantified using PCR amplification and Illumina sequencing. Lung tissues from four patients with bronchiectasis collected during the same period were used as controls. The RNA sequencing results were validated using immunohistochemistry (IHC) in another cohort (30 patients with NTM-PD and 15 patients with bronchiectasis). NTM-PD exhibited distinct gene expression patterns in T cells and macrophages. Gene set enrichment analysis revealed that pathways related to antigen presentation and processing were upregulated in NTM-PD, particularly in macrophages. Macrophages were more prevalent and the expression of genes associated with the M1 phenotype (CD40 and CD80) was significantly elevated. Although macrophages were activated in the NTM-PD group T cell activity was unaltered. Notably, expression of the costimulatory molecule CD28 was decreased in NTM-PD. IHC analysis showed that T cells expressing Foxp3 or TIM-3, which facilitate the regulatory functions of T cells, were increased. From these, NTM-PD exhibits distinct immunologic signatures characterized by the activation of macrophages without T cell activation.
2024-08-06 | GSE273714 | GEO
Project description:Immunologic features of nontuberculous mycobacterial pulmonary disease based on spatially resolved whole transcriptomes
| PRJNA1142902 | ENA