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Dive into the research topics where Márcia Susana Nunes Silva is active.

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Featured researches published by Márcia Susana Nunes Silva.


Journal of Clinical Microbiology | 2003

Mutations in katG, inhA, and ahpC Genes of Brazilian Isoniazid-Resistant Isolates of Mycobacterium tuberculosis

Márcia Susana Nunes Silva; Simone Gonçalves Senna; Marta Osório Ribeiro; Andréia Rosane de Moura Valim; Maria Alice da Silva Telles; Afrânio Lineu Kritski; Glenn P. Morlock; Robert C. Cooksey; Arnaldo Zaha; Maria Lucia Rosa Rossetti

ABSTRACT The presence of mutations in specific regions of the katG, inhA, and ahpC genes was analyzed with 69 Mycobacterium tuberculosis isoniazid-resistant isolates from three Brazilian states. Point mutations in codon 315 of the katG gene were observed in 87.1, 60.9, and 60% of the isolates from Rio Grande do Sul, Rio de Janeiro, and São Paulo, respectively. Mutations in the inhA gene were identified only in one isolate from RJ State, and the ahpC promoter region revealed mutations in distinct positions in 12.9, 21.7, and 6.7% of the isolates from RS, RJ and SP, respectively.


BMC Microbiology | 2009

Correlations of mutations in katG, oxyR-ahpC and inhA genes and in vitro susceptibility in Mycobacterium tuberculosis clinical strains segregated by spoligotype families from tuberculosis prevalent countries in South America.

Elis Regina Dalla Costa; Marta Osório Ribeiro; Márcia Susana Nunes Silva; Liane Silveira Arnold; Diana Rostirolla; Patrícia Izquierdo Cafrune; Roger Espinoza; Moises Palaci; Maria Alice da Silva Telles; Viviana Ritacco; Philip Noel Suffys; Maria Luiza Lopes; Creuza Campelo; Silvana Spindola de Miranda; Kristin Kremer; Pedro Eduardo Almeida da Silva; Leila de Souza Fonseca; John L. Ho; Afranio Lineu Kritski; Maria Lucia Rosa Rossetti

BackgroundMutations associated with resistance to rifampin or streptomycin have been reported for W/Beijing and Latin American Mediterranean (LAM) strain families of Mycobacterium tuberculosis. A few studies with limited sample sizes have separately evaluated mutations in katG, ahpC and inhA genes that are associated with isoniazid (INH) resistance. Increasing prevalence of INH resistance, especially in high tuberculosis (TB) prevalent countries is worsening the burden of TB control programs, since similar transmission rates are noted for INH susceptible and resistant M. tuberculosis strains.ResultsWe, therefore, conducted a comprehensive evaluation of INH resistant M. tuberculosis strains (n = 224) from three South American countries with high burden of drug resistant TB to characterize mutations in katG, ahpC and inhA gene loci and correlate with minimal inhibitory concentrations (MIC) levels and spoligotype strain family. Mutations in katG were observed in 181 (80.8%) of the isolates of which 178 (98.3%) was contributed by the katG S315T mutation. Additional mutations seen included oxyR-ahpC; inhA regulatory region and inhA structural gene. The S315T katG mutation was significantly more likely to be associated with MIC for INH ≥2 μg/mL. The S315T katG mutation was also more frequent in Haarlem family strains than LAM (n = 81) and T strain families.ConclusionOur data suggests that genetic screening for the S315T katG mutation may provide rapid information for anti-TB regimen selection, epidemiological monitoring of INH resistance and, possibly, to track transmission of INH resistant strains.


Revista De Saude Publica | 2002

Tuberculose resistente: revisão molecular

Maria Lucia Rosa Rossetti; Andréia Rosane de Moura Valim; Márcia Susana Nunes Silva; Vívian de F.S. Rodrigues

Progress to understanding the basis of resistance to antituberculous drugs has allowed molecular tests for detection of drug-resistant tuberculosis to be developed. Drug-resistant tuberculosis poses a threat to tuberculosis control programs. It is necessary thus to know drug susceptibilities of individual patients strain to provide the appropriate drug combinations. Molecular studies on the mechanism of action of antituberculous drugs have elucidated the genetic basis of drug resistance in M. tuberculosis. The mechanisms of drug resistance in tuberculosis are a result of chromosomal mutations in different genes of the bacteria. Upon drug exposure there is a selective pressure for such resistant mutants. Multidrug-resistant tuberculosis is a health problem of increasing significance for the whole global community. This paper reviews the molecular mechanisms associated with drug-resistance as well the new perspectives for detecting resistant isolates.


Memorias Do Instituto Oswaldo Cruz | 2011

Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

Candice Tosi Michelon; Franciele Rosso; Karen Barros Schmid; Rosa Dea Sperhacke; Martha Maria Oliveira; Afrânio Lineu Kritski; Leonides Rezende Jr; Elis Regina Dalla Costa; Andrezza Woloski Ribeiro; Patrícia Izquierdo Cafrune; Márcia Susana Nunes Silva; Daniele Kuhleis; Arnaldo Zaha; Maria Lucia Rosa Rossetti

Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis.


Diagnostic Microbiology and Infectious Disease | 2008

Phenotypic and genotypic characterization of drug-resistant Mycobacterium tuberculosis strains.

Wanessa Trindade Clemente; Stella Sala Soares Lima; Moises Palaci; Márcia Susana Nunes Silva; Vívian de F.S. Rodrigues; Elis Regina Dalla Costa; Lia Gonçalves Possuelo; Patrícia Izquierdo Cafrune; Fabíola Karla Ribeiro; Harisson M. Gomes; José Carlos Serufo

Of 142 pulmonary tuberculosis patients, 76 were considered high risk for the development of resistance, and 24 were confirmed as resistant strain carriers. Resistant isoniazid strains presented a high frequency of katG and ahpC mutations (90%) correlated with an MIC >4 microg/mL (94%). inhA mutations were not seen. rpoB mutations were identified in 78.6% of rifampicin-resistant strains, usually in codon 531 (72.7%), and 75% had an MIC >16 microg/mL. katG and rpoB mutations recognized 88.2% of multidrug-resistant strains and proved more efficient than the katG and rpoB mutations alone. Seventy percent of resistant pyrazinamide strains had pncA mutations between genes 136 and 188, 62.5% of them with an MIC >900 microg/mL. Pyrazinamidase inactivity was not an efficient resistance marker because 60% of pncA-mutated strains maintained enzymatic activity despite displaying good correlation with high resistance levels. Resistant ethambutol strains had embB mutations in codon 306, with MIC >16 microg/mL.


Mutation Research | 2011

Sequence and structural characterization of tbnat gene in isoniazid-resistant Mycobacterium tuberculosis: identification of new mutations.

Millene Borges Coelho; Elis Regina Dalla Costa; Sidra Vasconcellos; Natali Tejada Linck; Ricardo Martins Ramos; Hermes Luís Neubauer de Amorim; Philip Noel Suffys; Adalberto R. Santos; Pedro Eduardo Almeida da Silva; Daniela Fernandes Ramos; Márcia Susana Nunes Silva; Maria Lucia Rosa Rossetti

The present study was carried out to investigate the presence of polymorphism in the N-acetyltransferase gene of 41 clinical isolates of Mycobacterium tuberculosis, that were resistant to isoniazid (INH) with no mutations in the hot spots of the genes previously described to be involved in INH resistance (katG, inhA and ahpC). We observed single nucleotide polymorphisms (SNPs) in ten of these, including the G619A SNP in five isolates and an additional four so far un-described mutations in another five isolates. Among the latter SNPs, two were synonymous (C276T, n=1 and C375G, n=3), while two more non-synonymous SNPs were composed of C373A (Leu→Met) and T503G (Met→Arg) were observed in respectively one and two isolates. Molecular modeling and structural analysis based in a constructed full length 3D models of wild type TBNAT (TBNAT_H37Rv) and the isoforms (TBNAT_L125M and TBNAT_M168R) were also performed. The refined models show that, just as observed in human NATs, the carboxyl terminus extends deep within the folded enzyme, into close proximity to the buried catalytic triad. Analysis of tbnat that present non-synonymous mutations indicates that both substitutions are plausible to affect enzyme specificity or acetyl-CoA binding capacity. The results contribute to a better understanding of structure-function relationships of NATs. However, further investigation including INH-sensitive strains as a control group is needed to get better understanding of the possible role of these new mutations on tuberculosis control.


Memorias Do Instituto Oswaldo Cruz | 2011

Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay

Raquel de Abreu Maschmann; Márcia Susana Nunes Silva; Rosa Dea Sperhacke; Marta Osório Ribeiro; Philip Noel Suffys; Harrison Magdinier Gomes; Enrico Tortoli; Fiorella Marcelli; Arnaldo Zaha; Maria Lucia Rosa Rossetti

We used a colorimetric reverse dot blot hybridization (CRDH) assay to detect the presence of mutations in a specific region of the rpoB gene, associated with rifampin (RIF) resistance, in a panel of 156 DNAs extracted from 103 RIF-sensitive and 53 RIF-resistant cultures of Mycobacterium tuberculosis. When compared with the antimicrobial susceptibility test (AST), the sensitivity and specificity of the CRDH were 92.3% and 98.1%, respectively. When compared with sequencing, the sensitivity and specificity of the CRDH were 90.6% and 100%, respectively. To evaluate the performance of the assay directly in clinical specimens, 30 samples from tuberculosis patients were used. For these samples, the results of the CRDH were 100% consistent with the results of the AST and sequencing. These results indicate that the rate of concordance of the CRDH is high when compared to conventional methods and sequencing data. The CRDH can be successfully applied when a rapid test is required for the identification of RIF resistance in M. tuberculosis.


PLOS ONE | 2015

Hepatitis B Virus Genotype D Isolates Circulating in Chapecó, Southern Brazil, Originate from Italy

Carolina de Souza Gusatti; Cintia Costi; Maria Laura Halon; Tarciana Grandi; Arlete Ferrari Rech Medeiros; Claudia Silva; Selma A. Gomes; Márcia Susana Nunes Silva; Christian Niel; Maria Lucia Rosa Rossetti

Hepatitis B virus genotype A1 (HBV/A1), of African origin, is the most prevalent genotype in Brazil, while HBV/F predominates in the other South American countries. However, HBV/D is the most common in the three states of southern Brazil, where ‘islands’ of elevated prevalence, as Chapecó and other cities, have been described. In this study, 202 HBV chronic carriers attending in 2013 the viral hepatitis ambulatory of Chapecó, were investigated. In comparison with previous studies performed in the same ambulatory, a rapid aging of the HBV infected population was observed (mean age of the newly diagnosed patients increasing from 29.9 ± 10.3 years in 1996 to 44.4 ± 13.3 years in 2013), probably due to a singular vaccination schedule at Chapecó that included not only children but also adolescents. Phylogenetic and BLAST analyses (S region) classified 91 HBV isolates into genotypes A (n = 3) and D (n = 88). The majority of HBV/D isolates were closely related to D3 sequences. To understand the reasons for the absence or near absence of genotypes A and F, and how HBV/D was introduced in the south of Brazil, HBV/D infected patients were inquired about their genealogical and geographical origins. Forty-three (52%) patients have their four grandparents of Italian origin, vs. seven (8%) who have their four grandparents of Brazilian origin. At all, 65 out of 83 (78%) patients had at least one grandparent originating from Italy. Taking into consideration the fact that Italy is one of the few countries where subgenotype D3 is predominant, the results strongly suggested that HBV/D was introduced in Brazil through Italian immigration which culminated between 1870 and 1920.


Memorias Do Instituto Oswaldo Cruz | 2009

In house colorimetric reverse hybridisation assay for detection of the mutation most frequently associated with resistance to isoniazid in Mycobacterium tuberculosis

Raquel de Abreu Maschmann; Márcia Susana Nunes Silva; Elis Regina Dalla Costa; Marta Osório Ribeiro; Franciele Rosso; Philip Noel Suffys; Enrico Tortoli; Fiorella Marcelli; Arnaldo Zaha; Maria Lucia Rosa Rossetti

Mutations in the katG gene have been identified and correlated with isoniazid (INH) resistance in Mycobacterium tuberculosis isolates. The mutation AGC-->ACC (Ser-->Thr) at katG315 has been reported to be the most frequent and is associated with transmission and multidrug resistance. Rapid detection of this mutation could therefore improve the choice of an adequate anti-tuberculosis regimen, the epidemiological monitoring of INH resistance and, possibly, the tracking of transmission of resistant strains. An in house reverse hybridisation assay was designed in our laboratory and evaluated with 180 isolates of M. tuberculosis. It could successfully characterise the katG315 mutation in 100% of the samples as compared to DNA sequencing. The test is efficient and is a promising alternative for the rapid identification of INH resistance in regions with a high prevalence of katG315 mutants.


Journal of Medical Virology | 2016

Association between cytokine gene polymorphisms and outcome of hepatitis B virus infection in southern Brazil

Carolina de Souza Gusatti; Cintia Costi; Rúbia Marília de Medeiros; Maria Laura Halon; Tarciana Grandi; Arlete Ferrari Rech Medeiros; Claudia Silva; Rodrigo Rodenbusch; Márcia Susana Nunes Silva; Christian Niel; Maria Lucia Rosa Rossetti

A number of studies have demonstrated associations between cytokine gene polymorphisms and outcome of hepatitis B virus (HBV) infection. However, no general consensus has been reached, possibly due to differences between ethnic groups. In this study, 345 individuals living in southern Brazil, including 196 chronic HBV carriers and 149 subjects who had spontaneously recovered from acute infection, were enrolled to evaluate the influence of cytokine gene polymorphisms on the outcome of HBV infection. Most participants were of European descent. Genotyping of IL2‐330 G/T, IL4‐589C/T, IL6‐174 G/C, IL10‐592C/A, IL10‐1082 A/G, IL17A‐197 G/A, IL17A‐692 T/C, TNF‐α‐238 G/A, and TNF‐α‐308 G/A single nucleotide polymorphisms was performed by using the minisequencing (single base extension) method. By multivariable analysis, a statistically significant association was found between genotypic profile AA + GA in TNF‐α‐308 and chronic HBV infection (OR, 1.82; 95%CI, 1.01–3.27; P = 0.046). In southern Brazil, the carriers of the −308A allele in the TNF‐α gene promoter have a moderately higher risk of becoming chronic carriers in case of HBV infection. In addition, patients with chronic active hepatitis B (n = 60) exhibited a decreased frequency (3.3%) of the TNF‐238A allele when compared to that (14.8%) found among asymptomatic HBV carriers (n = 136), suggesting that this could be a protective factor against liver injury (OR, 0.17; 95%CI, 0.04–0.076; P = 0.023). J. Med. Virol. 88:1759–1766, 2016.

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Arnaldo Zaha

Universidade Federal do Rio Grande do Sul

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Elis Regina Dalla Costa

Federal University of Rio de Janeiro

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Marta Osório Ribeiro

Universidade Federal do Rio Grande do Sul

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Afrânio Lineu Kritski

Federal University of Rio de Janeiro

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Cristiana Alves Martins

Universidade Luterana do Brasil

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Graziele Lima Bello

Universidade Luterana do Brasil

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Moises Palaci

Universidade Federal do Espírito Santo

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