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Rapid identification and differentiation of Mycobacterium avium subspecies paratuberculosis types by use of real-time PCR and high-resolution melt analysis of the MAP1506 locus.

ABSTRACT: High-resolution melt (HRM) analysis can identify sequence polymorphisms by comparing the melting curves of amplicons generated by real-time PCR amplification. We describe the application of this technique to identify Mycobacterium avium subspecies paratuberculosis types I, II, and III. The HRM approach was based on type-specific nucleotide sequences in MAP1506, a member of the PPE (proline-proline-glutamic acid) gene family.

SUBMITTER: Castellanos E

PROVIDER: S-EPMC2849625 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature

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Rapid identification and differentiation of Mycobacterium avium subspecies paratuberculosis types by use of real-time PCR and high-resolution melt analysis of the MAP1506 locus.

Castellanos Elena E Aranaz Alicia A De Buck Jeroen J

Journal of clinical microbiology 20100203 4

High-resolution melt (HRM) analysis can identify sequence polymorphisms by comparing the melting curves of amplicons generated by real-time PCR amplification. We describe the application of this technique to identify Mycobacterium avium subspecies paratuberculosis types I, II, and III. The HRM approach was based on type-specific nucleotide sequences in MAP1506, a member of the PPE (proline-proline-glutamic acid) gene family. ...[more]

PMID: 20129970

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Molecular identification of Mycobacterium avium subsp. silvaticum by duplex high-resolution melt analysis and subspecies-specific real-time PCR.

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New PCR systems to confirm real-time PCR detection of Mycobacterium avium subsp. paratuberculosis.

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Mycobacterium avium subspecies paratuberculosis

Project description:Mycobacterium avium subspecies paratuberculosis Genome sequencing and assembly

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Characterization of the inflammatory phenotype of Mycobacterium avium subspecies paratuberculosis

Project description:Understanding the pathogenic mechanisms and host responses to Johne's disease, caused by Mycboacterium avium subspecies paratuberculosis (MAP), is complicated by a number of challenges of the disease progression and lack of ex vivo models to study the disease. We describe novel cell culture passage model which mimics the course of infection in vivo. Using this model we determined that cell culture passaged MAP develops an inflammatory phenotype initiating higher levels of pro-inflammatory cytokines and chemokines compared to levels initiated by non-passaged MAP. To verify that the MAP phenotypes were changing, we conducted transcriptome analysis of each population during the passage model and identified a change in gene expression, specifically and upregulation of genes involved in the biosynthesis of lipid components within the cell. Lipidomic analysis showed dramatically different lipid profiles between the 2 populations. These transcriptome changes were validated by the identification of high levels of upregulated transcripts from MAP-infected cattle ileal tissue. In total, this study described a novel model used to show that MAP changes phenotype during intracellular passage, thus changing its lipid profile and triggering an inflammatory response in the host leading to the severe clinical phage of Johne's disease.

2014-11-01 | GSE62566 | GEO

Mycobacterium avium Subspecies paratuberculosis: Human Exposure through Environmental and Domestic Aerosols.

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Neglected Facts on Mycobacterium Avium Subspecies Paratuberculosis and Type 1 Diabetes.

Project description:Civilization factors are responsible for the increasing of human exposure to mycobacteria from environment, water, and food during the last few decades. Urbanization, lifestyle changes and new technologies in the animal and plant industry are involved in frequent contact of people with mycobacteria. Type 1 diabetes is a multifactorial polygenic disease; its origin is conditioned by the mutual interaction of genetic and other factors. The environmental factors and certain pathogenetic pathways are shared by some immune mediated chronic inflammatory and autoimmune diseases, which are associated with triggers originating mainly from Mycobacterium avium subspecies paratuberculosis, an intestinal pathogen which persists in the environment. Type 1 diabetes and some other chronic inflammatory diseases thus pose the global health problem which could be mitigated by measures aimed to decrease the human exposure to this neglected zoonotic mycobacterium.

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High prevalence of viable Mycobacterium avium subspecies paratuberculosis in Crohn's disease.

Project description:AimTo examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis (MAP) in patients with inflammatory bowel disease [Crohn's disease (CD) and ulcerative colitis (UC)].MethodsThirty patients with CD (15 with at least one NOD2/CARD15 mutation), 29 with UC, and 10 with no inflammatory bowel disease (IBD). were tested for MAP by polymerase chain reaction (specific IS900 fragment) and blood culture.ResultsMAP DNA was detected in all original blood samples and 8-wk blood cultures (CD, UC and non-IBD). Positive MAP DNA status was confirmed by dot blot assays. All 69 cultures were negative by acid-fast Ziehl-Neelsen staining. Viable MAP, in spheroplast form, was isolated from the 18-mo blood cultures of all 30 CD patients, one UC patient, and none of the non-IBD controls. No association was found between positive MAP cultures and use of immunosuppressive drugs or CD-associated single nucleotide polymorphisms.ConclusionMAP is widely present in our area and MAP DNA can be recovered from the blood of CD, UC and non-IBD patients. However, MAP spheroplasts were only found in CD patients.

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| S-EPMC7157240 | biostudies-literature

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