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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

ABSTRACT: Diversity of nontuberculous mycobacterial infections in Kuwait

PROVIDER: PRJEB29664 | ENA |

REPOSITORIES: ENA

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Diversity of Nontuberculous Mycobacteria in Kuwait: Rapid Identification and Differentiation of Mycobacterium Species by Multiplex PCR, INNO-LiPA Mycobacteria v2 Assay and PCR Sequencing of rDNA.

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

Nontuberculous mycobacteria

Project description:Nontuberculous mycobacteria Metagenome

| PRJNA779755 | ENA

Gene signature profiles in respiratory epithelium infected with nontuberculous mycobacteria

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

Nontuberculous mycobacteria (NTM)

Project description:Genome Sequences and Assembly of Clinical and Environmental Isolates of Nontuberculous Mycobacteria

| PRJNA944390 | ENA

Mycobacterial communities in pulmonary NTM

Project description:Profiling Mycobacterial Communities in Pulmonary Nontuberculous Mycobacterial Disease

| PRJEB22153 | ENA

Immunologic features of nontuberculous mycobacterial pulmonary disease based on spatially resolved whole transcriptomes

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

Immunologic features of nontuberculous mycobacterial pulmonary disease based on spatially resolved whole transcriptomes

Project description:Immunologic features of nontuberculous mycobacterial pulmonary disease based on spatially resolved whole transcriptomes

| PRJNA1142902 | ENA

RNAseq to understand the host transcriptome during nontuberculous mycobacterial lung infection

Project description:We sought to elucidate the differentially expressed host factors in bronchoalveolar lavage cells (BALc) from nontuberculous mycobacteria (NTM)-infected subjects by studying the host transcriptome in during infection. A heatmap was created from the normalized expression of selected genes that showed statistically significant differential expression between NTM-infected and uninfected subjects. Genes with similar expression patterns were grouped and the most highly expressed genes are shown. One of the most differentially expressed gene in the NTM-infected group was the CFD (complement factor D) gene. Of note, a statistically significant three-fold increase in expression of the CAMP gene, encoding the LL-37 peptide was observed in the NTM-infected subject group (padj=0.05).

2018-09-30 | GSE103852 | GEO

Nontuberculous Mycobacteria by Metagenomic Next-Generation Sequencing

Project description:Nontuberculous Mycobacteria by Metagenomic Next-Generation Sequencing

| PRJEB43518 | ENA

Multiplex real-time PCR assay and melting curve analysis for identifying Mycobacterium tuberculosis complex and nontuberculous mycobacteria.

Project description:A multiplex real-time PCR assay and melting curve analysis for identifying 23 mycobacterial species was developed and evaluated using 77 reference strains and 369 clinical isolates. Concordant results were obtained for all 189 (100%) isolates of the Mycobacterium tuberculosis complex and 169 (93.9%) isolates of nontuberculous mycobacteria. Our results showed that this multiplex real-time PCR assay is an effective tool for the mycobacterial identification from cultures.

| S-EPMC3264165 | biostudies-literature

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