Resumo


A tuberculose pulmonar é uma doença infecciosa grave, que afeta aproximadamente 14 milhões de pessoas em todo o mundo, tendo 87 mil novos casos em 2009 no Brasil, país considerado com alta carga da doença. Os principais sintomas da tuberculose pulmonar são: tosse, escarro - por vezes sanguinolento, - dor torácica, fraqueza, perda de peso, febre e sudorese noturna. Estima-se que, durante um ano, um indivíduo com a forma pulmonar bacilífera poderá infectar, em média, de 10 a 15 pessoas. É uma doença curável na maioria dos casos novos, desde que o esquema terapêutico seja seguido e os pacientes sejam portadores de cepas de M. tuberculosis sensíveis aos fármacos anti-TB. A identificação do paciente sintomático respiratório é a principal estratégia para o controle da tuberculose.

Attachments:
Download this file (1509_BRATS_16.pdf)1509_BRATS_16.pdf[ ]539 kB
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Tavares RC, Salgado J, Moreira VB, Ferreira MA, Mello FC, Leung JW, Fonseca Lde S, Spallek R, Singh M, Saad MH.


Abstract

Several antigens of Mycobacterium tuberculosis have been identified and specificity to one or multiple antigens could determine the distinction between protective and pathogenic host reaction. Therefore T cell immune response to combinations 38 kDa/CFP-10, 38 kDa/MPT-64, ESAT-6/MPT-64 and ESAT-6/CFP-10 (each related to a single protein of Mycobacterium tuberculosis) in individuals from tuberculosis endemic areas have been examined. ELISA was used to detect IFN-gamma production in PBMC priming with single proteins and combinations in a panel of 105 individuals: 38 tuberculosis patients (6 untreated and 32 treated) and 67 healthy controls with tuberculin skin test positive or negative (TST). Brazilian TB patients highly recognized ESAT-6 (66%), but combinations improved response in the following order: ESAT-6/MPT-64 (89%) > ESAT-6/CFP-10 (73%) > 38 kDa/CFP-10 (70%), the last combination showing the highest specificity (TST(/) = 42% and TST(-) = 83%). Average IFN-gamma production in TB patients was signifi-cantly higher for 38 kDa/CFP-10 (P = 0.012) and 38 kDa/MPT-64 (P <0.035), when compared to single antigens.
 
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Hebe Rodrigues Cavalcanti; Elizabeth Marques; Leila de Souza Fonseca; Maria Helena Féres Saad

ABSTRACT

Double repetitive element (DRE) PCR amplification is a simple Mycobacterium tuberculosis typing method, however amplification failure or poor resolution of bands commit its efficacy. In order to verify if whether or not these features could be minimized by improving DNA extraction procedures or Taq polymerise quality, DRE-PCR was performed on 24 M. tuberculosis DNA samples extracted by heat-shock, mechanical and enzymatic methods applying conventional and hot start Taq pol. We demonstrated that when dealing with the Mycobacterium tuberculosis DRE-PCR typing method, Taq pol of better quality might be more important to improve amplification than the DNA extraction method.


Key words: Mycobacterium tuberculosis, PCR, Taq hot start, DRE-PCR

Attachments:
Download this file (1322_DNA_Extraction_Methods.pdf)1322_DNA_Extraction_Methods.pdf[ ]112 kB

Mayra Arias, Fernanda C. Q. Mello, Ada Pavón, Anna Grazia Marsico, Carlos Alvarado-Gálvez, Senia Rosales, Carlos Leonardo Carvalho Pessôa, Melly Pérez, Monica K. Andrade, Afranio L. Kritski, Leila S. Fonseca, Richard E. Chaisson, Michael E. Kimerling, and Susan E. Dorman


Abstract


Background. There is an urgent need for low-cost methods for rapid, accurate detection of Mycobacterium tuberculosis in clinical specimens. The microscopic-observation drug-susceptibility (MODS) assay is a relatively low-cost and simple liquid culture method that has been proposed for use in resource-limited environments.

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Eduardo Eustáquio de Souza Figueiredo; Flávia Galindo Silvestre; Wilma Neres Campos; Leone Vinícius Furlanetto; Luciana Medeiros; Walter Lilenbaum; Leila Sousa Fonseca; Joab Trajano Silva; Vânia Margaret Flosi Paschoalin


Isolates from suggestive bovine tuberculosis lesions were tested by a multiplex polymerase chain reaction (m-PCR) targeting for RvD1Rv2031c and IS6110 sequences, specific for M. bovis and Mycobacterium tuberculosis complex respectively. The m-PCR successfully identified as M. bovis 88.24% of the isolates.


Key words: Mycobacterium bovis, multiplex-PCR, bovine tuberculosis

Marcia Valéria B.S. Martinsae, Mônica Cristina B.S. Limab, Nadia C. Dupprea, Haroldo J. Matosc, John S. Spencerd, Patrick J. Brennand, Euzenir N. Sarnoa, Leila Fonsecae, Geraldo M.B. Pereiraab, Maria Cristina V. Pessolania

Summary


There are no reliable means for detecting subclinical mycobacterial infections. The recent sequencing of several mycobacterial genomes has now afforded new opportunities for the development of pathogen-specific diagnostic tests, critical in the context of leprosy and tuberculosis control. In the present study, we applied a multi-parametric flow cytometric analysis that allowed the investigation of T-cell functions in order to define immunological markers that measure previous exposure to mycobacteria. We compared the in vivo response to PPD, the gold standard skin test reagent for measuring previous exposure to Mycobacterium tuberculosis, with in vitro parameters of leukocyte activation in five PPD positive and five PPD negative healthy volunteers. PPD-stimulated peripheral leukocytes expressing CD4, CD69, cutaneous lymphocyte-associated antigen (CLA) and intracellular IFN-γ were enumerated in whole blood and compared with the size of in vivo PPD-induced induration and IFN-γ production levels as measured by ELISA in supernatants of PPD-stimulated peripheral blood mononuclear cells. The reactivity to the tuberculin skin test (TST) was associated with markedly increased frequencies of PPD-responsive activated (CD69+) and IFN-γ-producing CD4+T cells. Detection of PPD-specific IFN-γ producing leukocytes was restricted to CD4+T cells and a subset of these cells was shown to express the skin homing molecule CLA. Multiple linear regression modeling of responses to PPD showed the highest association between skin test indurations and frequencies of PPD-responsive IFN-γ-producing CD4+CD69+ T cells. Our data show that the in vitro enumeration of antigen-specific IFN-γ-producing CD4+ T cells can provide an alternative to the in vivo tuberculin test for the detection of latent Mycobacterium tuberculosis infection. Moreover, the measurement of these immunological parameters can be useful for the screening of new specific antigens defined by the genome sequence allowing selection of the best candidates for new diagnostics (including new skin tests), and vaccines for leprosy and tuberculosis.

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Giampaglia, C.M.S.; Martins, M.C.; Chimara, E.; Oliveira, R.S.; de Oliveira Vieira, G.B.; Marsico, A.G.; Mello, F.C.Q.; de Souza Fonseca, L.; Kritski, A.; da Silva Telles, M.A.

Abstract:

SETTING: Mycobacteria growth in media with the addition of inhibitory substances has been used in species identification. Growth of the Mycobacterium tuberculosis complex (MTC) is inhibited by ρ-nitrobenzoic acid (PNB), whereas non-tuberculous mycobacteria (NTM) are resistant.

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A Rede Brasileira de Pesquisa em Tuberculose (REDE-TB) é uma Organização Não Governamental (ONG) de direito privado sem fins lucrativos, preocupada em auxiliar no desenvolvimento não só de novos medicamentos, novas vacinas, novos testes diagnósticos e novas estratégias de controle de TB, mas também na validação dessas inovações tecnológicas, antes de sua comercialização no país e/ou de sua implementação nos Programa de Controle de TB no País.


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