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      Susceptibilidad de M. tuberculosis a drogas antituberculosas, determinada por dos técnicas, en el estado Sucre, Venezuela Translated title: usceptibility of M. tuberculosis to antituberculosis drugs as determined by two methods, in Sucre state, Venezuela

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          Abstract

          El objetivo de este trabajo fue evaluar la resistencia a isoniacida (INH), rifampicina (RIF), estreptomicina (STR) y etambutol (EMB) de 59 cepas de Mycobacterium tuberculosis, aisladas en el período agosto 2005-diciembre 2006, en el estado Sucre, Venezuela, empleando el método de proporciones de Canetti y de nitrato reductasa. Se encontró 6,3% de resistencia primaria y 14,3% de adquirida. Una cepa fue considerada MDR, al presentar resistencia a RIF e INH. Se comparó la prueba de nitrato reductasa con el método de las proporciones, encontrándose 100% de concordancia entre los resultados de los dos métodos para INH, RIF y EMB, y 95,65% para STR. Además, la prueba nitrato reductasa produjo resultados en 10 a 14 días, comparado con 42 días para el método de proporciones, por lo que la primera se postula como una alternativa muy valiosa para acortar el tiempo de respuesta en la valoración de la susceptibilidad de M. tuberculosis. La secuencia del gen rpoB en la cepa resistente a RIF demostró la presencia de una mutación no descrita anteriormente en la región hipervariable de 81 pares de bases, donde se ha reportado el mayor número de mutaciones de cepas resistentes a RIF. Esta mutación produjo un cambio en el codón 456 de TCG > CAG. Al comparar nuestros resultados con los hallados en el último estudio de prevalencia de resistencia realizado en el estado, se demuestra una disminución en la circulación de cepas resistentes en la zona de estudio

          Translated abstract

          The objective of this study was to evaluate the resistance to isoniazid (INH), rifampicin (RIF), streptomycin (STR) and ethambutol (EMB), with the Canetti’s proportions method (PM) and the nitrate reductase assay (NRA) of 59 clinical strains of Mycobacterium tuberculosis, isolated in the period of august 2005 to december 2006, in Sucre state, Venezuela. Primary and acquired drug resistance was 6.3% and 14.3%, respectively. Only one strain was found to be multidrug resistant (MDR). The overall agreement between the NRA and PM was 100% for INH, RIF and EMB, and 96% for STR. The time to obtain results was 10 to 14 days for the NRA, compared to 42 days for the PM. The NRA was easy to perform and therefore represents a useful tool for rapid and accurate determination of drug-resistant M. tuberculosis. The sequence of the rpoB gene of the RIF resistant strain demonstrated a never described mutation (change in the codon 456; TCG > CAG) in the hypervariable region of 81 base pairs where most of the mutations of the RIF resistant strains have been reported. Comparison of our results with those of the last resistance prevalence study carried out in the years 1998-1999, shows a decrease in the studied area

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          Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis.

          Control of tuberculosis is threatened by widespread emergence of drug resistance in Mycobacterium tuberculosis. Understanding the molecular basis of resistance might lead to development of novel rapid methods for diagnosing drug resistance. We set out to determine the molecular basis of resistance to rifampicin, a major component of multidrug regimens used for treating tuberculosis. Resistance to rifampicin involves alterations of RNA polymerase. The gene that encodes the RNA polymerase subunit beta (rpoB) was cloned. Sequence information from this gene was used to design primers for direct amplification and sequencing of a 411 bp rpoB fragment from 122 isolates of M tuberculosis. Mutations involving 8 conserved aminoacids were identified in 64 of 66 rifampicin-resistant isolates of diverse geographical origin, but in none of 56 sensitive isolates. All mutations were clustered within a region of 23 aminoacids. Thus, substitution of a limited number of highly conserved aminoacids encoded by the rpoB gene appears to be the molecular mechanism responsible for "single step" high-level resistance to rifampicin in M tuberculosis. This information was used to develop a strategy (polymerase chain reaction-single-strand conformation polymorphism) that allowed efficient detection of all known rifampicin-resistant mutants. These findings provide the basis for rapid detection of rifampicin resistance, a marker of multidrug-resistant tuberculosis.
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            Treatment of 171 patients with pulmonary tuberculosis resistant to isoniazid and rifampin.

            The frequency of infection with multidrug-resistant Mycobacterium tuberculosis is increasing. We reviewed the clinical courses of 171 patients with pulmonary disease due to M. tuberculosis resistant to rifampin and isoniazid who were referred to our hospital between 1973 and 1983. The patients' records were analyzed retrospectively. Their regimens were selected individually and preferably included three medications that they had not been given previously and to which the strain was fully susceptible. The 171 patients (median age, 46 years) had previously received a median of six drugs and shed bacilli that were resistant to a median of six drugs. Thus, their regimens were frequently not optimal. Of 134 patients with sufficient follow-up data, 87 (65 percent) responded to chemotherapy (as indicated by negative sputum cultures for at least three consecutive months); 47 patients (35 percent) had no response, as shown by continually positive cultures. The median stay in the hospital was more than seven months. In a multivariate analysis, an unfavorable response was significantly associated with a greater number of drugs received before the current course of therapy (odds ratio, 4.0; 95 percent confidence interval, 1.6 to 9.9; P < 0.001) and with male sex (odds ratio, 2.5; 95 percent confidence interval, 1.1 to 6.2; P < 0.03). Twelve of the patients with responses subsequently had relapses. The overall response rate was 56 percent over a mean period of 51 months. Of the 171 patients, 63 (37 percent) died, and 37 of these deaths were attributed to tuberculosis. For patients with pulmonary tuberculosis that is resistant to rifampin and isoniazid, even the best available treatment is often unsuccessful. Only about half of such patients eventually have negative sputum cultures despite carefully selected regimens administered for extended periods. Failure to control this resistant infection is associated with high mortality and ominous implications for the public health.
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              Analysis for a limited number of gene codons can predict drug resistance of Mycobacterium tuberculosis in a high-incidence community.

              Correct and rapid diagnosis is essential in the management of multidrug-resistant tuberculosis (MDR-TB). In this population-based study of 61 patients with drug-resistant tuberculosis, we evaluated the frequency of mutations and compared the performance of genotypic (mutation analysis by dot blot hybridization) and phenotypic (indirect proportion method) drug resistance tests. Three selected codons (rpoB531, rpoB526, and katG315) allowed identification of 90% of MDR-TB cases. Ninety percent of rifampin, streptomycin, and ethambutol resistance and 75% of isoniazid resistance were detected by screening for six codons: rpoB531, rpoB526, rrs-513, rpsL43, embB306, and katG315. The performance (reproducibility, sensitivity, and specificity) of the genotypic method was superior to that of the routine phenotypic method, with the exception of sensitivity for isoniazid resistance. A commercialized molecular genetic test for a limited number of target loci might be a good alternative for a drug resistance screening test in the context of an MDR "DOTS-plus" strategy.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                ic
                Investigación Clínica
                Invest. clín
                Universidad del Zulia (Maracaibo )
                0535-5133
                December 2010
                : 51
                : 4
                : 445-455
                Affiliations
                [1 ] Universidad de Oriente Venezuela
                [2 ] Hospital Vargas
                [3 ] IVIC
                [4 ] Hospital Dr. Julio Rodríguez Venezuela
                Article
                S0535-51332010000400002
                5d2a8165-1d03-4bce-a719-cf6d20eab812

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Venezuela

                Self URI (journal page): http://www.scielo.org.ve/scielo.php?script=sci_serial&pid=0535-5133&lng=en
                Categories
                MEDICINE, RESEARCH & EXPERIMENTAL

                Medicine
                Mycobacterium tuberculosis,resistance,multidrug-resistant,resistance assay,rpoB gene,resistencia,pruebas de resistencia,Gen rpoB

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