%0 Journal Article
%T Detecci車n de la expresi車n g谷nica in vivo de Mycobacterium tuberculosis durante la tuberculosis pulmonar activa Mycobacterium tuberculosis in vivo-expressed genes detection during active pulmonary tuberculosis
%A Alejandra Otazo M
%A Ingrid Guti谷rrez S
%A V赤ctor Acevedo F
%A Carlos Calder車n A
%J Revista Chilena de Enfermedades Respiratorias
%D 2012
%I Sociedad Chilena de Enfermedades Respiratorias
%X El estudio de la expresi車n g谷nica de Mycobacterium tuberculosis ha involucrado la experimentaci車n "in vitro ", "ex vivo " e "in vivo " (modelos animales), pero a迆n sin el 谷xito esperado. Proponemos que revelar los factores clave de la tuberculosis humana requiere investigar la expresi車n g谷nica de M. tuberculosis dentro del ser humano ("in vivo "). Para ello, aislamos el mRNA total de M. tuberculosis, desde muestras cl赤nicas respiratorias de pacientes con diagn車stico de tuberculosis pulmonar; posteriormente, sintetizamos el dscDNA y lo analizamos mediante RT-PCR cualitativo. Detectamos la expresi車n de la secuencia de inserci車n IS6110 y de los genes "housekeeping " 16SrRNA y sigA en M. tuberculosis creciendo in vivo (tuberculosis pulmonar) as赤 como cultivado in vitro. La expresi車n de los genes mprA y mprB, que codifican el sistema de transducci車n de se ales MprAB, s車lo se detect車 en M. tuberculosis crecido in vitro. Con nuestros resultados damos el primer paso hacia la implementaci車n de un m谷todo no invasivo para el estudio del transcriptoma de M. tuberculosis, dentro de su 迆nico hospedero natural, con el fin de analizar la regulaci車n "in vivo" de los determinantes gen谷ticos requeridos para su virulencia y patog谷nesis. Mycobacterium tuberculosis gene expression studies have involved "in vitro", "ex vivo" and "in vivo" experiments (animal models), but without the expected success. We propose that key features of human tuberculosis could be discovered by studying the M. tuberculosis gene expression within the human host. Therefore, we isolated totalM. tuberculosis mRNA from human clinical respiratory specimens of patients diagnosed with pulmonary tuberculosis; after this, we synthesized the dscDNA and tested it by qualitative RT-PCR assays. We detected the expression of IS6110 insertion sequence and of the "housekeeping" genes 16SrRNA andsigA in M. tuberculosis grown in vivo (pulmonary tuberculosis) as well as grown in vitro M. tuberculosis. mprA and mprB genes expression, which code the MprAB signal transduction system, were only detected in M. tuberculosis grown in vitro. Our results provide the first step towards a non invasive methodfor the study of the transcriptome of M. tuberculosis within its native host, to analyze "in vivo" regulation of the genetic determinants required for virulence and pathogenesis.
%K Mycobacterium tuberculosis
%K expresi車n in vivo
%K mRNA
%K tuberculosis pulmonar
%K Mycobacterium tuberculosis
%K in vivo expression
%K mRNA
%K pulmonary tuberculosis
%U http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-73482012000400004