Journal: medRxiv

Article Title: Direct detection and quantification of Mycobacterium tuberculosis from clinical samples by high-resolution melt qPCR

doi: 10.64898/2026.03.07.26347851

Figure Lengend Snippet: (A) Amplification plot of H37Rv replicates targeting the RD9 gene. (B) Corresponding melt curve showing a specific melting temperature (Tm) of ∼75°C. H37Rv DNA was serially diluted from 10 6 to 10 1 copies per reaction and analysed in triplicate. (C) Amplification plot obtained after 40 cycles for the dilutions, including the no-template control (NTC). (D) Corresponding melt curve showing a Tm of 73.7 ± 0.12°C. (E) Standard curve generated from the 10-fold serial dilutions, plotting cycle threshold (Ct) versus log copy number per reaction.

Article Snippet: Using the M. tuberculosis H37Rv (ATCC #25618DQ) reference strain, we performed asymmetric PCR with Vent (exo-) DNA polymerase (New England Biolabs).

Techniques: Amplification, Control, Generated

Journal: BMC Infectious Diseases

Article Title: Development and evaluation of an in-house single step loop-mediated isothermal amplification (SS-LAMP) assay for the detection of Mycobacterium tuberculosis complex in sputum samples from Moroccan patients

doi: 10.1186/s12879-016-1864-9

Figure Lengend Snippet: Nucleotide sequences of the SS-LAMP primers targeting the IS6110 sequence of MTBC strains

Article Snippet: SS-LAMP primers have been also tested on some MTBC strains DNA ( M. tuberculosis H37Rv, ATCC 25618D-2; M. tuberculosis H37Ra, ATCC 25177D-5 ; M. bovis, ATCC-BAA- 935D-2 ) and non-MTBC strains DNA ( M. avium, ATCC-BAA-968D-5 ; M. kansasii, ATCC 12478; M. intracellulare, ATCC 35769; M. abscessus, ATCC 19977D-5; M. gordonae, ATCC 14470D-5; M. smegmatis, ATCC 700084D-5; Escherichia coli, ATCC 8739; Pseudomonas aeruginosa, ATCC 9027; Staphylococcus aureus, ATCC 6538; Candida albicans, ATCC 10231; Aspergillus niger, ATCC 16888 ).

Techniques: Sequencing

Journal: Genome Medicine

Article Title: The variability and reproducibility of whole genome sequencing technology for detecting resistance to anti-tuberculous drugs

doi: 10.1186/s13073-016-0385-x

Figure Lengend Snippet: Study samples (DNA extracted from culture isolates) and their susceptibility to anti-tuberculosis drugs

Article Snippet: All clinical samples and the reference strain H37Rv (ATCC 25618D-9, Lot # 60986340) were prepared by inoculating a single colony into Middlebrook 7H9 broth supplemented with 10% OADC (Becton Dickinson) (see Table for list).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Small Molecule-directed Immunotherapy against Recurrent Infection by Mycobacterium tuberculosis *

doi: 10.1074/jbc.M114.558098

Figure Lengend Snippet: Stat-6 −/− CD4-TGFβRIIDN animals are resistant to M. tuberculosis infection. A , wild type BALB/c, STAT-6 −/− CD4-TGFβRIIDN, and STAT-6 −/− T-bet −/− CD4-TGFβRIIDN mice were infected with low dose (∼110 cfu) H37Rv through the aerosol route. Bacterial burdens were determined in the organs and at the time points indicated. Data represent mean ± S.D. values of four mice per group per time point, and the experiment was repeated twice. B , histological photomicrographs (×40) of lung sections (6 μm) of wild type BALB/c, STAT-6 −/− CD4-TGFβRIIDN, and STAT-6 −/− T-bet −/− CD4-TGFβRIIDN mice at 60 days after infection, stained with hematoxylin and eosin. C , ELISPOT assay to show the cytokine expression in CD4 + T cells in lung and spleen of M. tuberculosis -infected wild type BALB/c, STAT-6 −/− CD4-TGFβRIIDN, and STAT-6 −/− T-bet −/− CD4-TGFβRIIDN mice. D , prevalence of CD4 + CD25 + FoxP3 + T cells in BALB/c, STAT-6 −/− CD4-TGFβRIIDN, and STAT-6 −/− T-bet −/− CD4-TGFβRIIDN mice was determined by FACS analysis. E , wild type BALB/c and T-bet −/− mice were infected with low dose (∼110 cfu) H37Rv through the aerosol route. Bacterial burdens were determined in the organs and at the time points indicated. Data represent mean ± S.D. values of four mice per group per time point, and the experiment was repeated twice. F , photomicrographs (×40) of lung sections (6 μm) of T-bet −/− mice at 60 days after infection, stained with hematoxylin and eosin. G , cytokine expression in CD4 + T cells of lung and spleen from M. tuberculosis infected wild type BALB/c and T-bet −/− mice was determined by ELISPOT assay. H , prevalence of CD4 + CD25 + FoxP3 + T cells in pooled T cells obtained from lung and spleen of infected wild type BALB/c and T-bet −/− mice. Significant differences between the groups were determined by analysis of variance followed by Tukey's multiple comparison tests. Asterisk indicates statistical significance, p ≤ 0.05.

Article Snippet: M. tuberculosis H37Rv strain was grown in Middlebrook 7H9 broth (BD Biosciences, Sparks, MD) containing 0.02% Tween 80 to mid-log phase at 37 °C for 3 weeks, aliquoted, and frozen at −80 °C until use.

Techniques: Infection, Aerosol, Staining, Enzyme-linked Immunospot, Expressing, Comparison

Journal: The Journal of Biological Chemistry

Article Title: Small Molecule-directed Immunotherapy against Recurrent Infection by Mycobacterium tuberculosis *

doi: 10.1074/jbc.M114.558098

Figure Lengend Snippet: Stat-6 −/− and CD4-TGFβRIIDN animals are partially resistant to M. tuberculosis infection. A , wild type BALB/c, STAT-6 −/− , and STAT-6 −/− T-bet −/− mice were infected with low dose (∼110 cfu) H37Rv through the aerosol route. Bacterial burdens were determined in the organs and at the time points indicated. Data represent mean ± S.D. values of four mice per group per time point, and the experiment was repeated twice. B , histological photomicrographs (×40) of lung sections (6 μm) of STAT-6 −/− and STAT-6 −/− T-bet −/− mice at 60 days after infection. C , ELISPOT assay of cytokine expression in CD4 + T cells of lung and spleen from M. tuberculosis -infected wild type BALB/c, STAT-6 −/− , and STAT-6 −/− T-bet −/− mice. D , prevalence of CD4 + CD25 + FoxP3 + T cells was determined by FACS analysis. E , wild type BALB/c, CD4-TGFβRIIDN, and T-bet −/− CD4-TGFβRIIDN mice were infected with low dose (∼110 cfu) H37Rv through the aerosol route. Bacterial burdens were determined in the organs and at the time points indicated. Data represent mean ± S.D. values of four mice per group per time point, and the experiment was repeated twice. F , Histological photomicrographs (×40) of lung sections (6 μm) of CD4-TGFβRIIDN, and T-bet −/− CD4-TGFβRIIDN mice at 60 days after infection. G , ELISPOT assay of cytokine expression in CD4 + T cells of lung and spleen from M. tuberculosis -infected wild type BALB/c, CD4-TGFβRIIDN, and T-bet −/− CD4-TGFβRIIDN mice. H , prevalence of CD4 + CD25 + FoxP3 + T cells in BALB/c, CD4-TGFβRIIDN, and T-bet −/− CD4-TGFβRIIDN mice was determined by FACS analysis.

Article Snippet: M. tuberculosis H37Rv strain was grown in Middlebrook 7H9 broth (BD Biosciences, Sparks, MD) containing 0.02% Tween 80 to mid-log phase at 37 °C for 3 weeks, aliquoted, and frozen at −80 °C until use.

Techniques: Infection, Aerosol, Enzyme-linked Immunospot, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Small Molecule-directed Immunotherapy against Recurrent Infection by Mycobacterium tuberculosis *

doi: 10.1074/jbc.M114.558098

Figure Lengend Snippet: Simultaneous inhibition of Th2 cytokines and TGF-β protects against M. tuberculosis infection. Wild type BALB/c and mutant mice were infected (110 cfu/mice) through the aerosol route with M. tuberculosis (H37Rv). These mice were injected intraperitoneally with D4476 (a TGFβRI inhibitor) and/or suplatast tosylate (a Th2 inhibitor) at 32 nmol/g of body weight starting from the 1st day until the 44th day after aerosol challenge. At different time points (15, 30, 45 days), six mice from each group were sacrificed, and bacterial loads (cfu) were determined in lungs ( A ), spleen, and liver ( B ). C , histological photomicrographs (×10) of lung sections (6 μm) at 60 days after infection of the indicated mice, stained with hematoxylin and eosin. Arrows indicate granulomatic regions. Significant differences between the groups were determined by analysis of variance followed by Tukey's multiple comparison tests. Asterisk indicates statistical significance, p ≤ 0.05.

Article Snippet: M. tuberculosis H37Rv strain was grown in Middlebrook 7H9 broth (BD Biosciences, Sparks, MD) containing 0.02% Tween 80 to mid-log phase at 37 °C for 3 weeks, aliquoted, and frozen at −80 °C until use.

Techniques: Inhibition, Infection, Mutagenesis, Aerosol, Injection, Staining, Comparison