scholarly journals Identifying Mixed Mycobacterium tuberculosis Infection and Laboratory Cross-Contamination during Mycobacterial Sequencing Programs

2018 ◽  
Vol 56 (11) ◽  
Author(s):  
David H. Wyllie ◽  
Esther Robinson ◽  
Tim Peto ◽  
Derrick W. Crook ◽  
Adebisi Ajileye ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Process Development ◽  
Diagnostic Process ◽  
Cross Contamination ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type ◽  
The Public ◽  
Next Generation Sequencing Ngs ◽  
Mycobacterial Growth ◽  
Defining Sets

ABSTRACT The detection of laboratory cross-contamination and mixed tuberculosis infections is an important goal of clinical mycobacteriology laboratories. The objective of this study was to develop a method to detect mixtures of different Mycobacterium tuberculosis lineages in laboratories performing mycobacterial next-generation sequencing (NGS). The setting was the Public Health England National Mycobacteriology Laboratory Birmingham, which performs Illumina sequencing on DNA extracted from positive mycobacterial growth indicator tubes. We analyzed 4,156 samples yielding M. tuberculosis from 663 MiSeq runs, which were obtained during development and production use of a diagnostic process using NGS. The counts of the most common (major) variant and all other variants (nonmajor variants) were determined from reads mapping to positions defining M. tuberculosis lineages. Expected variation was estimated during process development. For each sample, we determined the nonmajor variant proportions at 55 sets of lineage-defining positions. The nonmajor variant proportion in the two most mixed lineage-defining sets (F2 metric) was compared with that of the 47 least-mixed lineage-defining sets (F47 metric). The following three patterns were observed: (i) not mixed by either metric; (ii) high F47 metric, suggesting mixtures of multiple lineages; and (iii) samples compatible with mixtures of two lineages, detected by differential F2 metric elevations relative to F47. Pattern ii was observed in batches, with similar patterns in the M. tuberculosis H37Rv control present in each run, and is likely to reflect cross-contamination. During production, the proportions of samples in the patterns were 97%, 2.8%, and 0.001%, respectively. The F2 and F47 metrics described could be used for laboratory process control in laboratories sequencing M. tuberculosis genomes.

scholarly journals Identifying mixed Mycobacterium tuberculosis infection and laboratory cross-contamination during Mycobacterial sequencing programs

2018 ◽  
Author(s):  
David H Wyllie ◽  
Esther Robinson ◽  
Tim Peto ◽  
Derrick W Crook ◽  
Adebisi Ajileye ◽  
...  
Keyword(s):  
Process Development ◽  
Tuberculosis Infection ◽  
Diagnostic Process ◽  
Cross Contamination ◽  
Mycobacterium Tuberculosis Infection ◽  
Next Generation Sequencing Ngs ◽  
Mycobacterial Growth ◽  
Indicator Tubes ◽  
Generation Sequencing ◽  
Defining Sets

ABSTRACTIntroductionDetecting laboratory cross-contamination and mixed tuberculosis infection are important goals of clinical Mycobacteriology laboratories.ObjectivesTo develop a method detecting mixtures of different M. tuberculosis lineages in laboratories performing Mycobacterial next generation sequencing (NGS).SettingPublic Health England National Mycobacteriology Laboratory Birmingham, which performs Illumina sequencing on DNA extracted from positive Mycobacterial Growth Indicator tubes.MethodsWe analysed 4,156 samples yielding M. tuberculosis from 663 MiSeq runs, obtained during development and production use of a diagnostic process using NGS. Counts of the most common (major) variant, and all other variants (non-major variants) were determined from reads mapping to positions defining M. tuberculosis lineages. Expected variation was estimated during process development.ResultsFor each sample we determined the non-major variant proportions at 55 sets of lineage defining positions. The non-major variant proportion in the two most mixed lineage defining sets (F2 metric) was compared with that in the 47 least mixed lineage defining sets (F47 metric). Three patterns were observed: (i) not mixed by either metric, (ii) high F47 metric suggesting mixtures of multiple lineages, and (iii) samples compatible with mixtures of two lineages, detected by differential F2 metric elevation relative to F47. Pattern (ii) was observed in batches, with similar patterns in the H37Rv control present in each run, and is likely to reflect cross-contamination. During production, the proportions of samples in each pattern were 97%, 2.8%, and 0.001%, respectively.ConclusionThe F2 and F47 metrics described could be used for laboratory process control in laboratories sequencing M. tuberculosis.

scholarly journals Neither Primary nor Memory Immunity to Mycobacterium tuberculosis Infection Is Compromised in Mice with Chronic Enteric Helminth Infection

2015 ◽  
Vol 83 (3) ◽  
pp. 1217-1223 ◽  
Author(s):  
Wasiulla Rafi ◽  
Kamlesh Bhatt ◽  
William C. Gause ◽  
Padmini Salgame
Keyword(s):  
Mycobacterium Tuberculosis ◽  
T Regulatory Cells ◽  
Helminth Infection ◽  
Treg Cells ◽  
Helminth Species ◽  
Nippostrongylus Brasiliensis ◽  
T Regulatory Cell ◽  
Chronic Infections ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type

Previously we had reported thatNippostrongylus brasiliensis, a helminth with a lung migratory phase, affected host resistance againstMycobacterium tuberculosisinfection through the induction of alternatively activated (M2) macrophages. Several helminth species do not have an obligatory lung migratory phase but establish chronic infections in the host that include potent immune downregulatory effects, in part mediated through induction of a FoxP3+T regulatory cell (Treg) response. Treg cells exhibit duality in their functions in host defense againstM. tuberculosisinfection since their depletion leads to enhanced priming of T cells in the lymph nodes and attendant improved control ofM. tuberculosisinfection, while their presence in the lung granuloma protects against excessive inflammation.Heligmosomoides polygyrusis a strictly murine enteric nematode that induces a strong FoxP3 Treg response in the host. Therefore, in this study we investigated whether host immunity toM. tuberculosisinfection would be modulated in mice with chronicH. polygyrusinfection. We report that neither primary nor memory immunity conferred byMycobacterium bovisBCG vaccination was affected in mice with chronic enteric helminth infection, despite a systemic increase in FoxP3+T regulatory cells. The findings indicate that anti-M. tuberculosisimmunity is not similarly affected by all helminth species and highlight the need to consider this inequality in human coinfection studies.

scholarly journals Disparate Effects of Metformin on Mycobacterium tuberculosis Infection in Diabetic and Nondiabetic Mice

2020 ◽  
Vol 65 (1) ◽  
pp. e01422-20
Author(s):  
Harindra D. Sathkumara ◽  
Karyna Hansen ◽  
Socorro Miranda-Hernandez ◽  
Brenda Govan ◽  
Catherine M. Rush ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Mycobacterium Tuberculosis ◽  
Disease Burden ◽  
Low Dose ◽  
Diabetic Mice ◽  
Antidiabetic Drug ◽  
Mycobacterium Tuberculosis Infection ◽  
Therapeutic Concentration ◽  
Content Type

ABSTRACTComorbid type 2 diabetes poses a great challenge to the global control of tuberculosis. Here, we assessed the efficacy of metformin (MET), an antidiabetic drug, in mice infected with a very low dose of Mycobacterium tuberculosis. In contrast to diabetic mice, infected nondiabetic mice that received the same therapeutic concentration of MET presented with significantly higher disease burden. This warrants further studies to investigate the disparate efficacy of MET against tuberculosis in diabetic and nondiabetic individuals.

scholarly journals Clonal Complexity in Mycobacterium tuberculosis Can Hamper Diagnostic Procedures

2017 ◽  
Vol 55 (5) ◽  
pp. 1388-1395 ◽  
Author(s):  
Laura Pérez-Lago ◽  
Marta Herranz ◽  
Yurena Navarro ◽  
María Jesús Ruiz Serrano ◽  
Pilar Miralles ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Risk ◽  
Mixed Infection ◽  
Diagnostic Procedures ◽  
Hiv Positive ◽  
Mixed Infections ◽  
Cross Contamination ◽  
Content Type ◽  
High Burden ◽  
Susceptibility Patterns

ABSTRACT Clonal complexity is increasingly accepted in Mycobacterium tuberculosis infection, including mixed infections by ≥2 strains, which usually occur in settings with a high burden of tuberculosis and/or a high risk of overexposure to infected patients. Mixed infections can hamper diagnostic procedures; obtaining an accurate antibiogram is difficult when the susceptibility patterns of the strains differ. Here, we show how mixed infections can also prove challenging for other diagnostic procedures, even outside settings where mixed infections are traditionally expected. We show how an unnoticed mixed infection in an HIV-positive patient diagnosed in Madrid, Spain, with differences in the representativeness of the coinfecting strains in different sputum samples, markedly complicated the resolution of a laboratory cross-contamination false positivity alert.

scholarly journals Efficient 5-OP-RU-Induced Enrichment of Mucosa-Associated Invariant T Cells in the Murine Lung Does Not Enhance Control of Aerosol Mycobacterium tuberculosis Infection

2020 ◽  
Vol 89 (1) ◽  
pp. e00524-20 ◽  
Author(s):  
Charles Kyriakos Vorkas ◽  
Olivier Levy ◽  
Miroslav Skular ◽  
Kelin Li ◽  
Jeffrey Aubé ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Bacterial Infections ◽  
Cell Activation ◽  
Bacterial Load ◽  
Cell Subset ◽  
Mycobacterium Tuberculosis Infection ◽  
Class I Mhc ◽  
Content Type ◽  
Mait Cells ◽  
Mait Cell

ABSTRACTMucosa-associated invariant T (MAIT) cells are an innate-like T cell subset in mammals that recognize microbial vitamin B metabolites presented by the evolutionarily conserved major histocompatibility complex class I (MHC I)-related molecule, MR1. Emerging data suggest that MAIT cells may be an attractive target for vaccine-induced protection against bacterial infections because of their rapid cytotoxic responses at mucosal services to a widely conserved bacterial ligand. In this study, we tested whether a MAIT cell priming strategy could protect against aerosol Mycobacterium tuberculosis infection in mice. Intranasal costimulation with the lipopeptide Toll-like receptor (TLR)2/6 agonist, Pam2Cys (P2C), and the synthetic MR1 ligand, 5-OP-RU, resulted in robust expansion of MAIT cells in the lung. Although MAIT cell priming significantly enhanced MAIT cell activation and expansion early after M. tuberculosis challenge, these MAIT cells did not restrict M. tuberculosis bacterial load. MAIT cells were depleted by the onset of the adaptive immune response, with decreased detection of granzyme B+ and gamma interferon (IFN-γ)+ MAIT cells relative to that in uninfected P2C/5-OP-RU-treated mice. Decreasing the infectious inoculum, varying the time between priming and aerosol infection, and testing MAIT cell priming in nitric oxide synthase 2 (NOS2)-deficient mice all failed to reveal an effect of P2C/5-OP-RU-induced MAIT cells on M. tuberculosis control. We conclude that intranasal MAIT cell priming in mice induces early MAIT cell activation and expansion after M. tuberculosis exposure, without attenuating M. tuberculosis growth, suggesting that MAIT cell enrichment in the lung is not sufficient to control M. tuberculosis infection.

scholarly journals Evaluation of Gamma Interferon Immune Response Elicited by the Newly Constructed PstS-1(285-374):CFP10 Fusion Protein To Detect Mycobacterium tuberculosis Infection

2014 ◽  
Vol 21 (4) ◽  
pp. 552-560 ◽  
Author(s):  
Leonardo Silva de Araujo ◽  
Fernanda Carvalho de Queiroz Mello ◽  
Nidai de Bárbara Moreira da Silva ◽  
Janaina Aparecida Medeiros Leung ◽  
Silvia Maria Almeida Machado ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Tuberculin Skin Test ◽  
Skin Test ◽  
Positive Tuberculin Skin Test ◽  
Gamma Interferon ◽  
Preventative Treatment ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type ◽  
Ifn Γ ◽  
First Time

ABSTRACTThe PstS1 antigen is highly immunogenic, principally when combined with CFP10 during both latent and active TB infection. In the present study, a selectedpstS1gene fragment was cloned, fused with CFP10, and expressed inEscherichia coli. The product [PstS-1(285-374):CFP10] was compared to the recombinant fused RD1 (region of deletion 1) protein (ESAT-6:CFP10) in detectingMycobacterium tuberculosisinfection in 108 recent contacts of pulmonary tuberculosis (TB) cases, considering a positive tuberculin skin test (TST) to be the baseline. The release of gamma interferon (IFN-γ) in 22-h whole-blood and 5-day lymphocyte stimulation assays primed with each antigen was determined. All contacts were clinically followed for up to 1 year, and 87% of the tuberculin skin test-positive (TSTpositive) patients accepted preventative treatment. Concerning the IFN-γ response to PstS-1(285-374):CFP10 in the 22-h and 5-day assays, a slight increase in contact-TSTpositivedetection was observed (23/54 and 26/54) compared to the level seen with the RD1 protein (18/54 and 24/54) whereas in the TSTnegativegroup, similarly lower numbers (≤5/48) of responders were achieved for both antigens, except for RD1 in the 5-day assay (8/48). By combining the IFN-γ responders to both antigens in the 5-day assays, slightly higher increases in positivity were found in the TSTpositive(32/54) and TSTnegative(10/48) groups. Two of 12 untreated TSTpositivecontacts progressed to active TB and were concordantly positive in all assays, except for one contact who lacked positivity in the RD1 5-day assay. We demonstrated for the first time that PstS-1(285-374):CFP10 slightly increased contact positivity and detection of active disease progression, suggesting its potential application as a TB infection marker.

scholarly journals An Atypical Manifestation of Mycobacterium tuberculosis Infection

2021 ◽  
Author(s):  
Iván Fernández Castro ◽  
María Jesús Isorno-Porto ◽  
Ignacio Novo-Veleiro ◽  
Clara Casar-Cocheteux ◽  
Lucía Barrera-López ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Imaging Techniques ◽  
Extrapulmonary Tuberculosis ◽  
Tuberculosis Infection ◽  
Diagnostic Process ◽  
Mycobacterium Tuberculosis Infection ◽  
The Past ◽  
Aortic Aneurysm And Dissection ◽  
Serious Disease ◽  
Atypical Manifestation

Introduction: Aortitis is seen in a wide variety of diseases. It was rarely found in the past but this is changing because of new imaging techniques. Case description: We present the case of a 45-year-old man who was found on thyroid ultrasound to have infrarenal aortitis and pathological lymphadenopathies in different locations. After an exhaustive diagnostic process, tuberculous aortitis, an infrequent manifestation of extrapulmonary tuberculosis, was diagnosed. The condition resolved after a 6-month course of antibiotics and a 6-week course of corticosteroids. Conclusion: Tuberculous aortitis is an atypical manifestation of Mycobacterium tuberculosis infection. The absence of typical symptoms and the difficulty of isolating the microorganism makes its diagnosis difficult. Therefore, clinical suspicion, microbiological tests and imaging are key for reaching the diagnosis and starting treatment for a serious disease that can cause aortic aneurysm and dissection.

scholarly journals Acute Inflammation Confers Enhanced Protection against Mycobacterium tuberculosis Infection in Mice

2021 ◽  
Author(s):  
Tucker J. Piergallini ◽  
Julia M. Scordo ◽  
Paula A. Pino ◽  
Larry S. Schlesinger ◽  
Jordi B. Torrelles ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Mycobacterium Tuberculosis ◽  
Causative Agent ◽  
Acute Inflammation ◽  
Causes Of Death ◽  
Tuberculosis Infection ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type ◽  
High Level ◽  
Tuberculosis Disease

Mycobacterium tuberculosis , the causative agent of tuberculosis disease, is estimated to infect one-fourth of the world’s population and is one of the leading causes of death due to an infectious disease worldwide. The high-level variability in tuberculosis disease responses in the human populace may be linked to immune processes related to inflammation.

GPR183 regulates interferons and bacterial growth during Mycobacterium tuberculosis infection: interaction with type 2 diabetes and TB disease severity

2020 ◽  
Author(s):  
Stacey Bartlett ◽  
Adrian Tandhyka Gemiarto ◽  
Minh Dao Ngo ◽  
Haressh Sajiir ◽  
Semira Hailu ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Mycobacterium Tuberculosis ◽  
Disease Severity ◽  
Mycobacterial Infection ◽  
Mycobacterium Tuberculosis Infection ◽  
Important Regulator ◽  
Chest X Ray ◽  
Mycobacterial Growth

AbstractOxidized cholesterols have emerged as important signaling molecules of immune function, but little is known about the role of these oxysterols during mycobacterial infections. We found that expression of the oxysterol-receptor GPR183 was reduced in blood from patients with tuberculosis (TB) and type 2 diabetes (T2D) compared to TB patients without T2D and was associated with TB disease severity on chest x-ray. GPR183 activation by 7α,25-hydroxycholesterol (7α,25-OHC) reduced growth of Mycobacterium tuberculosis (Mtb) and Mycobacterium bovis BCG in primary human monocytes, an effect abrogated by the GPR183 antagonist GSK682753. Growth inhibition was associated with reduced IFN-β and IL-10 expression and enhanced autophagy. Mice lacking GPR183 had significantly increased lung Mtb burden and dysregulated IFNs during early infection. Together, our data demonstrate that GPR183 is an important regulator of intracellular mycobacterial growth and interferons during mycobacterial infection.Graphical Abstract

scholarly journals The Endothelin System Has a Significant Role in the Pathogenesis and Progression of Mycobacterium tuberculosis Infection

2014 ◽  
Vol 82 (12) ◽  
pp. 5154-5165 ◽  
Author(s):  
Andre F. Correa ◽  
Alexandre M. Bailão ◽  
Izabela M. D. Bastos ◽  
Ian M. Orme ◽  
Célia M. A. Soares ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type ◽  
Host Interactions ◽  
Endothelin System ◽  
Etb Receptor ◽  
The Impact ◽  
The Relationship ◽  
Host Tissues

ABSTRACTTuberculosis (TB) remains a major global health problem, and although multiple studies have addressed the relationship betweenMycobacterium tuberculosisand the host on an immunological level, few studies have addressed the impact of host physiological responses. Proteases produced by bacteria have been associated with important alterations in the host tissues, and a limited number of these enzymes have been characterized in mycobacterial species.M. tuberculosisproduces a protease called Zmp1, which appears to be associated with virulence and has a putative action as an endothelin-converting enzyme. Endothelins are a family of vasoactive peptides, of which 3 distinct isoforms exist, and endothelin 1 (ET-1) is the most abundant and the best-characterized isoform. The aim of this work was to characterize the Zmp1 protease and evaluate its role in pathogenicity. Here, we have shown thatM. tuberculosisproduces and secretes an enzyme with ET-1 cleavage activity. These data demonstrate a possible role of Zmp1 for mycobacterium-host interactions and highlights its potential as a drug target. Moreover, the results suggest that endothelin pathways have a role in the pathogenesis ofM. tuberculosisinfections, and ETA or ETB receptor signaling can modulate the host response to the infection. We hypothesize that a balance between Zmp1 control of ET-1 levels and ETA/ETB signaling can allowM. tuberculosisadaptation and survival in the lung tissues.

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