A live, attenuated form of Mycobacterium bovis, bacillus Calmette–Guérin (BCG), is commonly used as intravesical immunotherapy for non-invasive urothelial bladder carcinoma. While complications are rare, dissemination can occur. A case of mycotic aortic aneurysm following BCG administration with recovery of Mycobacterium bovis in culture is reported. A review of the published experience with this problem is also presented.

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Background: In this present study we decided to consider the prevalence and distribution of Beijing family in the world using meta-analysis based on systematic review of articles published and relation with drug resistance, which will provide more detailed information to clearly overview the status of this family and transmission of TB. Methods: This study used the most available article published in literature database including PubMed, Science direct, Web of Science, Google Scholar, Biological abs, Iranmedex, and SID systematically reviewed prevalence of Beijing family. Data analyzed using meta-analysis with random effects models. Results: Final analyses included 264 samples that have been selected from 2811 studies. Overall Beijing family prevalence in world was estimated to be 33.2% (95% CI 31.4–35.2). Corresponding estimates by continent were Asia 44.7% (39.5–49.8), Europe 27.9% (25.6–30.1), Africa 12×5% (8.9–16.2), and America 8.9% (6.9–10.9). In all world regions, Beijing families were associated with drug resistance 81.37%. Conclusions: According to the results, prevalence of Beijing family in Asia is higher than similar studies in other parts of the world and this family is associated with drug resistance. Effective control program is needed in world to control the spread of drug resistance strains specially Beijing family.

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Objective: Neopterin is a sensitive marker for cell-mediated immune response. Because of this, the neopterin levels of body fluids show cell-mediated immune response in different infectious diseases which involve T cells and macrophages. The aim of this study was to determine the clinical importance of neopterin levels in patients with tuberculosis and compare with those levels of healthy subjects. Methods: Seventy patients with tuberculosis (46 newly diagnosed cases, 15 relapse cases, and 9 multidrug-resistant tuberculosis cases) and 18 healthy adult individuals were included in the study. Neopterin concentrations were measured by the ELISA method according to the protocol of the manufacturer. Chi-square test was used in statistical analysis; p≤0.05 was considered statistically significant. Results: Serum mean neopterin levels were 23.74±21.8nmol/L (median: 18.3) in newly diagnosed patients with pulmonary tuberculosis; 28.69±21.2nmol/L (median: 21.2) in relapse patients and 31.28±14nmol/L (median: 25.4) in multidrug-resistant tuberculosis cases, respectively. Serum mean neopterin levels were 4.03±5.12nmol/L (median: 5.1) in healthy subjects. The serum neopterin levels were found to be significantly higher in patients with tuberculosis than the control group. There was a statistically significant correlation between neopterin positivity (neopterin level ≥10nmol/L was accepted to be positive) and clinical symptoms of hemoptysis and weight loss. Besides statistically significant correlations between neopterin positivity and hemoglobin level, sedimentation rate, mean leukocyte count and radiological involvement (localized or diffuse) were determined. Conclusion: Serum neopterin levels can be used as a helper laboratory finding for the diagnosis of patients with tuberculosis. For this aim, further controlled studies are needed.

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Background and objective: The epidemiology of the forty percent of tuberculosis patients who present with disseminated and/or extrapulmonary disease is in need of further study. Further study of such dissemination using published data from international indices may provide data which assist with control of tuberculosis. Methods: For each clinical or epidemiologic factor studied, summary odds ratios and corresponding 95% confidence intervals were calculated showing associations between such factors and documented extrapulmonary dissemination of tuberculosis. Results: Eighteen studies fulfilled criteria for study of the clinical factors and nine for the cytokine studies. Significant factors associated with a greater risk of extrapulmonary dissemination were female gender (summary odds ratio, 1.92 (95% confidence intervals, 1.72–2.13), I-squared 86.9), age under 45 (1.37, 1.18–1.60, 63.7), and as well the absence of smoking, drinking and diabetes but not HIV infection (1.10, 0.91–1.32, 80.5). Among cytokines, the macrophage receptor protein P2X7 was associated most strongly associated with extrapulmonary dissemination of tuberculosis (2.28, 0.88–5.90, 92.9). Conclusion: Young age, female gender, and the macrophage purinergic receptor protein P2X7 were major factors associated with extrapulmonary dissemination of tuberculosis.

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Bacillus Calmette–Guérin (BCG) remains the only widely used vaccine against tuberculosis (TB). Consistent efficacy has been proved in infants but not in adults from developing countries. Epidemiological and experimental studies have pointed out that, prior exposure to prevailing environmental mycobacteria could be responsible for the poor efficacy of BCG as an anti-TB vaccine in adults living in developing countries. Sensitization by environmental mycobacteria may down-modulate the immunologic behavior of BCG on the one hand and may mask its efficacy on the other hand. Some of the important deciding factors for poor efficacy of BCG, due to exposure of the subjects to prevailing environmental mycobacteria, are thought to be (i) Life stage: neonatus versus adolescence; (ii) shared antigens between prevailing environmental mycobacteria and BCG; and (iii) generation of cross-reactive T-regulatory cells against environmental mycobacteria and BCG. In this communication, some novel strategies have been discussed for countering the down modulating impact of environmental mycobacteria towards performance of BCG as an anti-TB vaccine.

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Background: In earlier studies, it was shown that ex vivo Mycobacterium tuberculosis-infected type II alveolar epithelial cells generate de novo nitric oxide (NO), but the mycobactericidal quantity of NO was released only by stimulation of these cells with proinflammatory cytokines, i.e. IFN-γ, TNF-α and IL-1β. In the present communication, it was demonstrated that M. tuberculosis-infected/mycobacterial antigens stimulated cells utilize both, JAK-STAT and NF-κB pathways for the induction of inducible Nitric Oxide Synthase (iNOS) mRNA and NO production. Methods: Alveolar epithelial cell line A549 were either infected with M. tuberculosis or stimulated with M. tuberculosis components. Confocal microscopy, NO estimation and EMSA were performed on the infected/stimulated A549 cells. Results: Nuclear extracts prepared from M. tuberculosis infected A549 cells alone or stimulated with IFN-γ or a combination of three cytokines (IFN-γ, TNF-α and IL-1β) formed DNA protein complexes with probes from both −5.2kb region (specific for binding of STAT-1 protein) and −5.8kb region (specific for binding of both STAT-1 and NF-κB) of the iNOS promoter. However, TNF-α or IL-1β stimulated M. tuberculosis-infected A549 cells showed no protein DNA complexes with construct from −5.2kb region. Conclusions: This differential response indicated that TNF-α/IL-1β does not allow STAT-1 production or its translocation to nucleus in M. tuberculosis-infected A549 cells in the absence of IFN-γ. This differential signaling of iNOS induction in M. tuberculosis-infected alveolar epithelial cells by cytokines may be responsible for controlled production of NO intracellularly.

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Background: Species specific diagnosis of mycobacterial infection is crucial because treatment of infections caused by Mycobacterium tuberculosis (MTB) differs from that of non-tuberculous mycobacterial (NTM) species. The species identification used to be cumbersome and non-reproducible a decade ago. Objectives: Recently, some commercial tests have been made available to differentiate the MTB and NTM growths in culture media. Sensitivity and specificity of these tests was evaluated. Materials and methods: In this double blind study 572 clinical samples were cultured in an automated BACTEC-MGIT-960 system. A total of 147 (25.7%) samples were MGIT culture positive. These cultures were subjected to an in-house m-PCR (which amplifies hsp-65, esat-6 and ITS region for MAC), two commercial immune-chromatographic tests (ICTs) and phenotypic tests. Results: Of the 147 MGIT positive cultures, m-PCR was able to correctly identify MTB in 123 cultures and NTM in 24 which included 3 MAC isolates. m-PCR showed 100% agreement with two gold standard methods-the nitrate reductase assay and PNB tests-in correctly identifying MTB. Commercial strips were able to correctly identify MTB in 120 (97.5%) of 123 cultures, while 3 (2.5%) isolates were falsely identified as NTM. However, none of the growth negative spent medium gave false positive results in any of the tests. None of the commercial strips misidentified any of the 24 NTM as MTB; hence, specificity of these strips was 100%. Of the 2 IC test systems, both SD Bioline and BD TBc strip tests missed 2.5% of MTB isolates and misidentified these as NTM. Conclusion: The in-house m-PCR was found to be the most accurate and efficient tool for identifying the MTB, MAC and other NTMs.

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Tuberculosis (TB) is caused by Mycobacterium tuberculosis (MTB) and the disease has remained a major health problem in most of the developing countries, particularly after the emergence of multidrug-resistant TB (MDR-TB). The MDR-TB is an intriguing subject and very little is known about the in vivo processes which take place during the acquisition of MDR. This study describes a unique case of pulmonary TB (PTB) from which four sequential isolates of MTB could be isolated while the patient was on anti-tubercular treatment. The first baseline isolate was sensitive to all drugs, but the subsequent three isolates acquired resistance to multiple drugs and finally the patient died after 27 months post-diagnosis when his fourth isolate became resistant to isoniazid, rifampicin, ethambutol and kanamycin. All sequential cultures were identified as MTB using conventional and molecular methods, including 16s RNA sequencing and the spoligotyping. Spoligotyping followed by comparison with SITVITWEB database revealed that all the isolates belonged to the family of the Central Asian Strain Delhi (CAS1_Delhi, ST26) genotype, and no cross or mixed infections were observed. The drug resistance was further characterized at the molecular level by sequencing the target genes (katG, inhA, rpoB, embB, eis promoter region and rrs). The results revealed mutated alleles associated with resistance to the respective drugs. This unique case indicates that it is possible to isolate MTB during treatment if the strain is acquiring resistance. The data presented from four sequential isolates provides an insight into what sequential genetic and proteomic changes occur in the bacteria during the in vivo acquisition of MDR.

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Osteoarticular tuberculosis represents 1.7–2% of all tuberculosis (TB). The localization in the foot is rare and accounts for less than 10% of osteoarticular TB. The following report describes the case of a 7-year-old boy who presented with a gradually increasing inflammatory swelling over the lateral aspect of the right ankle. An X-ray of his right ankle showed an osteolytic image at the calcaneus. Diagnosis was confirmed by the presence of a tuberculoid granuloma with caseous necrosis on bone biopsy.

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Sporotrichoid tuberculosis is a rare form of cutaneous tuberculosis; it primarily affects children after a post-traumatic inoculation. The diagnosis is often difficult and based on a set of arguments; it should be considered in any sporotrichoid lesion, especially in tuberculosis endemic countries. The following describes a new case of Mycobacterium tuberculosis skin infection with an unusual sporotrichoid clinical appearance in a healthy woman, emphasizing the diagnostic difficulties with a review of literature.

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Background: Mycobacterium tuberculosis is known to slow down its transcriptional activity during dormancy. Hence, while using reporter strains, it is important to couple the reporter gene to a promoter that is strong and sensitive both in active and dormant M. tuberculosis. Since respiration is an indispensable process even in dormant bacteria, validation of the promoters of respiratory chain genes – type II NADH dehydrogenase (P_ndh) and adenosine triphosphate (ATP) synthase operon (P_atps) – of MTB was undertaken for this purpose. Methods: Putative promoter containing sequences were cloned upstream of a red fluorescent protein (RFP) gene. Mycobacterium smegmatis or M. tuberculosis carrying episomal constructs were validated for growth, fitness and fluorescence in different models in vitro and in vivo. Results: Either promoter can drive stable and strong expression of RFP in actively growing and dormant M. smegmatis in vitro without significantly affecting growth or viability. Fluorescence due to P_ndh and P_atps was significantly higher than P_hsp60. The fitness of M. tuberculosis H₃₇Rv counterparts was unaffected inside J774 macrophages. In immunocompetent mice, despite an initial attenuation in the lungs, both strains reached loads similar to wild type during chronic infection. In the spleen, the fluorescent strain counts were similar to wild type counts throughout. RFP fluorescence in tissue homogenates was more homogenous among mice due to P_ndh compared with P_atps. Conclusions: Coupling an appropriate reporter to the promoter of ndh-2 gene of M. tuberculosis can make the reporter expression respiration sensitive and thereby reliably detect both active and dormant populations of the reporter strain.

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A study was conducted at Rayagada district of Odisha, India, among smear-positive tuberculosis (TB) patients to determine the resistance pattern to first-line drugs. Sputum samples were collected from 405 new and 37 previously treated patients and were tested at Regional Medical Research Centre, Bhubaneswar. Resistance to any anti-tubercular drug was observed to be 5.2% among new cases and 16.1% among previously treated patients, while multidrug-resistant tuberculosis (MDR-TB) was found to be 0% in new and 8.1% in previously treated cases. Such a low level of resistance may be due to the limited use of TB drugs outside the ongoing program.

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Tuberculosis is a global health problem which has been compounded by the emergence and rapid spread of drug resistant strains. Phenotypic drug susceptibility testing of Mycobacterium tuberculosis usually requires homogenization of cultures using 3–5 mm glass beads. In resource limited settings, these important material may either not be readily available in the country as in our case requiring that one orders them from abroad or they may be too expensive. In both situations, this would impact on the usually lean budget. In our centre were we recently introduced tuberculosis culture and drug susceptibility testing using the Microscopic Observation Drug Susceptibility (MODS) technique, we successfully used glass fragments from a broken car windshield obtained from a mechanic workshop to homogenize solid cultures to prepare positive controls. All cultures homogenized with these local beads gave consistent MODS results. The challenge of the limited availability of resources for research in resource limited settings can be met by adapting available materials to achieve results.

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Tuberculosis is explicitly recognized as a major global public health problem. In Côte d'Ivoire, relapse cases represent 66.5% of patients eligible for retreatment according to the National Tuberculosis Control Program. This study objective was to detect multidrug-resistance tuberculosis among relapse cases. Patients were recruited in tuberculosis centers in routine. A standardized questioning was administrated. Two sputum samples were collected and transported at Institut Pasteur. Sputum samples were decontaminated by NALC method. The DNA extraction was realized with 500 μl of decontaminated sputum sample with smear-positive. MTBDRplus assay version 2.0 was performed according to the manufacturer's instruction. An internal quality control program with positive and negative controls was implemented for interpretation of results. In total 146 relapse cases with smear positive were studied. Out of selected patients, 130 had received the 2RHZE/4RH regimen and 16, the 2RHZES/1RHZE/5HRE. In group of relapse cases previously treated with 2RHZE/4RH regimen, 40 (31.3%, IC95%: [0.23; 0.39]) had punctual mutations at codon 526 in rpoB gene. Although, in patients under treated with 2RHZES/1RHZE/5HRE, a mutation in rpoB gene was identified in 12 of 16 sputum samples. Thirteen mutations conferring a resistance to Isoniazid were observed of which 9 in katG gene and 4 in katG and promoter region of inhA gene. The comparison (Chi-square with Yates correction) of resistance rates to Rifampin estimated showed a statistically significant difference. Conclusion: Use of a rapid method to detect drug-resistance in recurrent TB cases has permitted to identify patients eligible for first-line drugs or not.

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