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The current study evaluated Mycobacterium tuberculosis isolates from Rio de Janeiro, Brazil, for genomic deletions. One locus in our panel of PCR targets failed to amplify in ϳ30% of strains. A single novel long sequence polymorphism (>26.3 kb) was characterized and designated RD Rio . Homologous recombination between two similar protein-coding genes is proposed as the mechanism for deleting or modifying 10 genes, including two potentially immunogenic PPE proteins. The flanking regions of the RD Rio locus were identical in all strains bearing the deletion. Genetic testing by principal genetic group, spoligotyping, variable-number tandem repeats of mycobacterial interspersed repetitive units (MIRU-VNTR), and IS6110-based restriction fragment length polymorphism analysis cumulatively support the idea that RD Rio strains are derived from a common ancestor belonging solely to the Latin American-Mediterranean spoligotype family. The RD Rio lineage is therefore the predominant clade causing tuberculosis (TB) in Rio de Janeiro and, as indicated by genotypic clustering in MIRU-VNTR analysis, the most significant source of recent transmission. Limited retrospective reviews of bacteriological and patient records showed a lack of association with multidrug resistance or specific risk factors for TB. However, trends in the data did suggest that RD Rio strains may cause a form of TB with a distinct clinical presentation. Overall, the high prevalence of this genotype may be related to enhanced virulence, transmissibility, and/or specific adaptation to a Euro-Latin American host population. The identification of RD Rio strains outside of Brazil points to the ongoing intercontinental dissemination of this important genotype. Further studies are needed to determine the differential strain-specific features, pathobiology, and worldwide prevalence of RD Rio M. tuberculosis.Tuberculosis (TB) is a preventable and curable infectious disease that nonetheless remains a significant cause of morbidity and mortality in resource-poor nations (14). TB also threatens to reemerge in developed nations as a consequence of increased immigration, its synergy with the human immunodeficiency virus (HIV)/AIDS epidemic, and a deprioritization of TB control efforts in public health policy (64). The principal etiologic agent of human TB is Mycobacterium tuberculosis, but other species within the M. tuberculosis complex (MTC), such as "Mycobacterium canettii" (proposed name), Mycobacterium africanum, and Mycobacterium bovis, are also known to cause TB in humans (30). Although the MTC is considered a relatively homogenous taxon at the DNA sequence level, an increasing number of species-, lineage-, and strain-specific genetic variations have been revealed by the identification of multicopy repeat elements, targeted interrogations of specific genetic loci, and the comparison of whole-genome sequences of several MTC species and strains (21,24,31). These differences have been exploited as markers for epidemiological purposes and/or to assist in the identific...
Immune mediators associated with human tuberculosis (TB) remain poorly defined. This study quantified levels of lung immune mediator gene expression at the time of diagnosis and during anti-TB treatment using cells obtained by induced sputum. Upon comparison to patients with other infectious lung diseases and volunteers, active pulmonary TB cases expressed significantly higher levels of mediators that counteract Th1-type and innate immunity. Despite the concomitant heightened levels of Th1-type mediators, immune activation may be rendered ineffectual by high levels of intracellular (SOCS and IRAK-M) and extracellular (IL-10 and TGF-βRII, IL-1Rn, and IDO) immune suppressive mediators. These modulators are a direct response to Mycobacterium tuberculosis as, by day 30 of anti-TB treatment, many suppressive factors declined to that of controls whereas most Th1-type and innate immune mediators rose above pretreatment levels. Challenge of human immune cells with M. tuberculosis in vitro up-regulated these immune modulators as well. The observed low levels of NO synthase-2 produced by alveolar macrophages at TB diagnosis, along with the heightened amounts of suppressive mediators, support the conclusion that M. tuberculosis actively promotes down-modulatory mediators to counteract Th1-type and innate immunity as an immunopathological strategy. Our data highlight the potential application of immune mediators as surrogate markers for TB diagnosis or treatment response.
103 SNP RFLP, as compared to results obtained using a PCR method targeting a LAM-restricted IS6110 element, correctly identified 99.8% of LAM spoligotype strains. Together, these tests were more accurate than spoligotyping at categorizing strains with indefinable spoligotypes and segregated true LAM strains from those with convergent spoligotypes. The fact that RD Rio strains were identified worldwide highlights the importance of this LAM family sublineage and suggests that this strain is a global threat that should be specifically targeted by public health resources. Our provision of simple and robust molecular methods will assist the evaluation of the LAM family and the RD Rio sublineage.
Molecular genotyping has shown Mycobacterium tuberculosis lineages to be geographically restricted and associated with distinct ethnic populations. Whether tuberculosis (TB) caused by some M. tuberculosis lineages can present with a differential clinical spectrum is controversial because of very limited clinical data. We recently reported on the discovery of RD Rio M. tuberculosis, a Latin American-Mediterranean sublineage that is the predominant cause of TB in Rio de Janeiro, Brazil. To investigate the clinical attributes of TB caused by RD Rio strains, we studied a cohort of TB cases from Belo Horizonte, Brazil, in which clinical information recorded on a standardized questionnaire was collected at the time of microbiological testing. These patients were referred for culture and drug susceptibility testing because of the clinical suspicion of "complicated" TB, as demonstrated by high rates of multidrug resistance (12%) and cavitary TB (80%). We performed spoligotyping and RD Rio genotyping on the M. tuberculosis strains and analyzed the clinical data from these patients. RD Rio M. tuberculosis accounted for 37% of the total TB burden. Multivariate analysis found a significant association between TB caused by RD Rio strains and pulmonary cavitation and residence in Belo Horizonte. Since cavitary TB is associated with higher sputum bacillary load, our findings support the hypothesis that RD Rio M. tuberculosis is associated with a more "severe" disease as a strategy to increase transmission. Future studies are needed to confirm these observations and to better define the contribution of RD Rio M. tuberculosis to the global TB epidemic.Mycobacterium tuberculosis, the etiologic agent of tuberculosis (TB), is estimated to have infected one-third of the world's population and annually causes ϳ8 million new TB cases and Ͼ2 million deaths (16, 58). The challenges posed by TB have been further worsened by the emergence of multi-drugresistant and extensively drug-resistant M. tuberculosis strains.Molecular typing, based on genetic markers, permits the rapid detection and species level identification of mycobacteria within the M. tuberculosis complex (MTC), as well as provides useful tools for examining the transmission and evolution of these microorganisms (7,17,30,53). Genome-wide single nucleotide polymorphism (SNP) and deletion analyses have been used to organize the global M. tuberculosis population structure into overlapping phylogenies with major lineages that show distinct geographic distribution and that may be associated with specific host adaptation (1,21,23,24,27,28,50). Similar results have also been obtained with spoligotyping and IS6110-RFLP fingerprint analysis (18). The W/Beijing lineage, for example, is the predominant family in major SNP cluster II, accounts for ϳ10% of strains causing TB globally, and is localized mainly to Asia but has spread internationally. W/Beijing has been associated with outbreaks and multiple drug resistance (MDR) (4, 32) and also contributed significantly to the resurgence of...
Mycobacterium bovis is the causative agent of bovine tuberculosis, with a wide host range. Fifty human M. bovis isolates were typed using spoligotyping and variable number tandem repeats (VNTR). Fifteen of these spoligotypes have not yet been recorded in cattle. The predominant spoligotype in humans and cattle was subdivided by VNTR.
Background Genetic tracking of Mycobacterium tuberculosis is a cornerstone of tuberculosis (TB) control programs. The RDRio M. tuberculosis sublineage was previously associated with TB in Brazil. We investigated 3847 M. tuberculosis isolates and registry data from New York City (NYC) (2001–2005) to: 1) affirm the position of RDRio strains within the M. tuberculosis phylogenetic structure, 2) determine its prevalence, and 3) define transmission, demographic, and clinical characteristics associated with RDRio TB. Methods Isolates classified as RDRio or non-RDRio M. tuberculosis by multiplex PCR were further classified as clustered (≥2 isolates) or unique based primarily upon IS6110-RFLP patterns and lineage-specific cluster proportions were calculated. The secondary case rate of RDRio was compared with other prevalent M. tuberculosis lineages. Genotype data were merged with the data from the NYC TB Registry to assess demographic and clinical characteristics. Results RDRio strains were found to: 1) be restricted to the Latin American-Mediterranean family, 2) cause approximately 8% of TB cases in NYC, and 3) be associated with heightened transmission as shown by: i) a higher cluster proportion compared to other prevalent lineages, ii) a higher secondary case rate, and iii) cases in children. Furthermore, RDRio strains were significantly associated with US-born Black or Hispanic race, birth in Latin American and Caribbean countries, and isoniazid resistance. Conclusions The RDRio genotype is a single M. tuberculosis strain population that is emerging in NYC. The findings suggest that expanded RDRio case and exposure identification could be of benefit due to its association with heightened transmission.
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