OBJETIVO: Descrever a prevalência de tuberculose multirresistente (TBMR) em pacientes com tuberculose em uma importante cidade brasileira através do II Inquérito Nacional de Resistência aos Fármacos Antituberculose, assim como as características sociais, demográficas e clínicas desses pacientes. MÉTODOS: De 2006 a 2007, amostras clínicas de pacientes de três hospitais e das cinco unidades básicas de saúde participantes do inquérito realizado em Porto Alegre foram coletadas e submetidas ao teste de sensibilidade aos fármacos. A confirmação das espécies de micobactérias ocorreu por métodos bioquímicos. RESULTADOS: Foram incluídos 299 pacientes. Desses, 221 (73,9%) eram homens e 77 (27,3%) tinham história de tuberculose. A idade média foi de 36 anos. Dos 252 pacientes testados para HIV, 66 (26,2%) estavam infectados. A prevalência da TBMR na amostra geral foi de 4,7% (IC95%: 2,3-7,1); enquanto essa foi de 2,2% (IC95%: 0,3-4,2) nos pacientes virgens de tratamento e de 12,0% (IC 95%: 4,5-19,5) naqueles com história de tratamento antituberculose. A análise multivariada mostrou que história de tuberculose e maior tempo para o diagnóstico associaram-se a TBMR. CONCLUSÕES: Caso esses resultados sejam confirmados em outros estudos no Brasil, a história de tratamento antituberculose e o maior tempo para o diagnóstico poderão ser utilizados como preditores de TBMR.

OBJECTIVE: To describe the prevalence of multidrug-resistant tuberculosis (MDR-TB) among tuberculosis patients in a major Brazilian city, evaluated via the Second National Survey on Antituberculosis Drug Resistance, as well as the social, demographic, and clinical characteristics of those patients. METHODS: Clinical samples were collected from tuberculosis patients seen between 2006 to 2007 at three hospitals and five primary health care clinics participating in the survey in the city of Porto Alegre, Brazil. The samples were subjected to drug susceptibility testing. The species of mycobacteria was confirmed using biochemical methods. RESULTS: Of the 299 patients included, 221 (73.9%) were men and 77 (27.3%) had a history of tuberculosis. The mean age was 36 years. Of the 252 patients who underwent HIV testing, 66 (26.2%) tested positive. The prevalence of MDR-TB in the sample as a whole was 4.7% (95% CI: 2.3-7.1), whereas it was 2.2% (95% CI: 0.3-4.2) among the new cases of tuberculosis and 12.0% (95% CI: 4.5-19.5) among the patients with a history of tuberculosis treatment. The multivariate analysis showed that a history of tuberculosis and a longer time to diagnosis were both associated with MDR-TB. CONCLUSIONS: If our results are corroborated by other studies conducted in Brazil, a history of tuberculosis treatment and a longer time to diagnosis could be used as predictors of MDR-TB.

The occurrence of tuberculosis (TB) in prisons has been described as an alarming public health problem in many countries, especially in developing nations. The objective of this study was to conduct a survey among prisoners with TB respiratory symptoms in order to estimate the incidence of the disease, to analyze the drug susceptibility profile and genotype the isolates of Mycobacterium tuberculosis in the city of Charqueadas, southern of Brazil. The TB incidence was 55/1,900 inhabitants in the prison; this corresponds to an incidence of 3,789/100,000 inhabitants, with a prevalence of 72/1,900 (4,960/100,000 inhabitants). Drug susceptibility test was performed and, among the analyzed isolates, 85% were susceptible to all drugs tested and 15% were resistant to at least one drug, of which 89% were resistant only to isoniazid (INH) or in combination with another drug. The genotype classification of spoligotyping analysis showed that 40% of the isolates belong to LAM family, 22% to T family, 17.5% to Haarlem family, 12.5% to U family and 3% to X family. The shared international spoligotypes most frequently found were 729 (27%), 50 (9.5%), 42 (8%), 53 (8%) and 863 (8%). In conclusion, it was observed that TB in this specific population had been caused, mostly, by strains that have been transmitted in the last few years, as demonstrated by the large level of genotype clustering. In addition, it was found specific large clusters, which were not often found in the general population from the same period and in the same region.

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.

Tuberculosis is an increasingly important public health problem in developing countries. We studied 111 tuberculosis patients confirmed by mycobacteria isolation between 1998 and 2000 in Rio Grande, in south Brazil. One-hundred-thirteen pulmonary and extrapulmonary clinical materials were examined through culture by the Ogawa-Kudoh method and through smear examination by the Ziehl-Nielsen or Kinyoun methods. The identification of Mycobacterium tuberculosis was done through the usual phenotypical methods. The proportion method (indirect technique) was used to determine the resistance of isolates. The man/woman ratio was 2.6:1, 75% were white, and the group had a mean age of 39.7±12.7. Risk factors were present in 38.7% (34 men and 9 women, p=0.2), with a mean age of 35 (p=0.002); infection by HIV was the most frequent (28.8%), followed by alcoholism (16.2%) and injected drug use (15.3%). The frequency of M. tuberculosis isolates was 99.1%. Pulmonary disease occurred in 88.3% of the cases, extrapulmonary in 9.9% and 1.8% in both. There was an association between extrapulmonary tuberculosis and the presence of risk factors (p=0.0001). Resistance to isoniazid was found in 4% of the isolates and to isoniazid and rifampin in 2%, all being patients with some risk factors. The profile of tuberculosis in the population of this study followed the pattern described for developing regions of the world.

INTRODUÇÃO: As taxas de resistência aos fármacos constituem um dos pilares da avaliação dos programas de controle da tuberculose. A demora na obtenção dos resultados, conseqüência da metodologia convencional utilizada, faz com que haja a necessidade de avaliação de novos testes, mais rápidos e menos onerosos. OBJETIVO: Comparar técnicas fenotípicas rápidas para determinação do perfil de susceptibilidade de M. tuberculosis, utilizando indicadores de viabilidade celular, com o teste das proporções em Löwenstein-Jensen, padrão-ouro. MÉTODO: Foram utilizadas 166 cepas de M. tuberculosis com o perfil de susceptibilidade conhecido. A concentração mínima inibitória de cada fármaco foi determinada, em microplaca, utilizando-se meio líquido e os indicadores de oxi-redução, Alamar Blue® e brometo de tetrazolium. O ponto de corte entre a cepa sensível e a resistente foi estabelecido como concentração mínima inibitória maior ou igual a 0,2 mg /mL para isoniazida e 1,0 mg /mL para rifampicina. RESULTADOS: Houve concordância total entre os dois métodos de determinação da concentração mínima inibitória. Comparando os resultados dos testes com o padrão-ouro, obteve-se uma concordância de 95%, para isoniazida e rifampicina. O tempo para obtenção dos resultados foi de 7 dias, contrastando com os 28 dias pelo método convencional. CONCLUSÃO: Os testes para determinação da concentração mínima inibitória, em meio líquido, utilizando indicadores de oxi-redução, são rápidos e podem se utilizados como alternativa rápida na determinação de susceptibilidade de cepas de M. tuberculosis.

BACKGROUND: Knowledge of the rates of drug resistance is one of the pillars of tuberculosis control program evaluation. Data from low-resource countries are scarce and results are delayed due to the techniques employed. There is therefore an urgent need for evaluation of faster and less onerous testing methods. OBJECTIVE: To compare the performance of rapid colorimetric assays for phenotyping that employ oxidation-reduction indicators to determine the susceptibility profile of Mycobacterium tuberculosis with the gold-standard proportion method on Lowenstein-Jensen Medium. METHOD: We analyzed 166 M. tuberculosis strains of known susceptibility. Minimal inhibition concentrations for isoniazid and rifampicin were determined in microplates, using a liquid medium and Alamar Blue and tetrazolium bromide indicators. To measure agreement the Kappa value was used. Cutoff values between sensitive and resistant strains were defined as 0.2mg/mL and 1.0mg/mL for isoniazid and rifampicin, respectively. RESULTS: There was 100% concordance between Alamar Blue and tetrazolium bromide methods in the determination of minimal inhibition concentrations. Agreement between the colorimetric method and the Lowenstein-Jensen was 95% for isoniazid and rifampicin. Using the colorimetric method, results were obtained within 7 days, in contrast to the 28 days required for the conventional method. CONCLUSIONS: Assays to determine minimal inhibition concentrations in liquid medium and employing oxidation-reduction indicators proved to be rapid and inexpensive. This method has the potential to become a faster, alternative method for determining susceptibility of M. tuberculosis strains in developing countries.

INTRODUÇÃO: A tuberculose é uma doença antiga que ainda se mantém como um dos maiores males da humanidade no século XXI. Nas últimas décadas, o advento de novas tecnologias utilizando os conhecimentos de biologia molecular tem levado a um aumento na investigação da etiologia, detecção e epidemiologia da tuberculose. OBJETIVO: Avaliar o grau de similaridade entre as cepas de Mycobacterium tuberculosis provenientes do setor de tisiologia do Centro de Saúde Navegantes, de Porto Alegre (RS). MÉTODO: Foi realizado um estudo retrospectivo utilizando 55 amostras de escarro de pacientes atendidos ambulatorialmente no Centro de Saúde Navegantes para realização da técnica de RFLP. Os resultados obtidos pela genotipagem foram correlacionados com os dados gerados a partir da epidemiologia convencional. RESULTADOS: Trinta e nove isolados (70,9%) apresentaram padrão único, enquanto dezesseis isolados (29,1%) apresentaram padrões agrupáveis e formaram 8 clusters, com 2 pacientes em cada. Foi encontrada relação epidemiológica em 6 (37,5%) dos 16 pacientes em cluster. CONCLUSÃO: A associação adequada entre epidemiologia convencional e genotipagem de M. tuberculosis contribui para um melhor entendimento da dinâmica de transmissão da tuberculose mesmo quando o estudo é realizado em um único local.

BACKGROUND: Tuberculosis is an ancient disease, which still remains one of the major ills faced by mankind in the 21st century. In recent decades, new technologies employing the knowledge gained from molecular biology studies have allowed for more accurate detection of tuberculosis and increased investigation of the etiology and epidemiology of the disease. AIM: Evaluating the degree of similarity among strains of Mycobacterium tuberculosis provided by the Phthisiology Sector of Centro de Saúde Navegantes (Navegantes Health Clinic) in Porto Alegre, RS, Brazil. METHOD: A retrospective study was performed involving RFLP typing of 55 sputum samples from outpatients examined at the Centro de Saúde Navegantes. The results of the genotyping were correlated to the conventional epidemiology data. RESULTS: A single pattern was seen in 39 (70.9%) of the isolates, whereas 16 isolates (29.1%) showed clustering patterns and were grouped into 8 clusters of 2 patients each. An epidemiological link was found for 6 (37.5%) of the 16 patients in the clusters. CONCLUSION: The appropriate combination of conventional epidemiology and genotyping of M. tuberculosis contributes to a better understanding of the dynamics of tuberculosis transmission even when such a study is performed in a single, isolated health clinic.

INTRODUÇÃO: A sinergia entre a tuberculose e o vírus da imunodeficiência humana é responsável pelo aumento da morbi-mortalidade dos pacientes com AIDS. OBJETIVO: Delinear o perfil de pacientes com tuberculose e AIDS na cidade de Rio Grande (RS) relacionando dados demográficos, clínicos e laboratoriais. MÉTODO: A amostra foi constituída por todos os casos de tuberculose confirmados pelo isolamento de Mycobacterium tuberculosis ocorridos no Serviço de AIDS do Hospital Universitário/FURG entre setembro de 1997 e dezembro de 2000, em 31 pacientes reportados como casos definidos de AIDS. Foram examinados 33 materiais clínicos pulmonares e extrapulmonares através da cultura pelo método de Ogawa-Kudoh e da baciloscopia pelo Kinyoun. A identificação de M. tuberculosis foi feita pelos métodos fenotípicos usuais. Para determinação da resistência das cepas isoladas foi empregado o método das proporções. RESULTADOS: A média de idade foi de 33,8 ± 9,9 anos, com uma relação homem/mulher de 2,87:1. Eram brancos 80,7% dos pacientes. Todos os pacientes apresentavam manifestações clínicas gerais e/ou específicas de tuberculose no momento da suspeita diagnóstica. Em 20 deles foram constatados fatores de risco: uso de droga endovenosa, alcoolismo, desnutrição, encarceramento. A doença pulmonar ocorreu em 19 casos, a extrapulmonar em 10 e a associada em 2 deles. Entre aqueles com a forma extrapulmonar, predominou o comprometimento ganglionar. As 33 cepas isoladas foram identificadas como M. tuberculosis, e 28 mostraram sensibilidade à isoniazida e à rifampicina. CONCLUSÃO: A tuberculose nos pacientes com AIDS apresentou-se com manifestações clínicas variáveis, comprometendo homens e mulheres em condições sociais desfavoráveis, em plena fase produtiva de suas vidas.

BACKGROUND: Synergism between tuberculosis and HIV is responsible for the increased morbidity-mortality rate in AIDS patients. OBJECTIVE: To delineate the profile of patients with tuberculosis and AIDS in the city of Rio Grande by relating demographic, clinical and laboratory data. METHOD: The sample comprised all cases of tuberculosis defined by identification of Mycobacterium tuberculosis that occurred in the AIDS Service of the University Hospital/FURG between September, 1997 and December, 2000, which added to a total of 31 patients confirmed as definite cases of AIDS. Using the Ogawa-Kudoh culture method and the Kinyoun bacilloscopy, 33 clinical pulmonary and extrapulmonary specimens were analyzed. Identification of M. Tuberculosis was made by the usual phenotype methods. The method of proportions was chosen to establish resistance of isolated strains. RESULTS: The mean age was of 33.8±9.9 years, with a man/ woman ratio of 2.87:1 and 80.7% of Caucasians. All patients (n=31) exhibited overall or specific clinical manifestations of turberculosis at the time of suspicion diagnosis. In 20 of the cases risk factors were observed: use of injected drugs, alcoholism, malnutrition, imprisonment. Pulmonary disease occurred in 19 cases, extrapulmonary in 10 and the association of both in two. Lymph node commitment was more frequent among those with extrapulmonary disease. The isolated strains (33) were identified as M. Tuberculosis and 28 were tested and showed sensibility to Isoniazid and Rifampin. CONCLUSION: In AIDS patients, tuberculosis appeared with various clinical manifestations, jeopardizing both men and women of less favored social conditions while at a fully productive stage of their lives.

No presente estudo, nós reportamos os resultados de uma análise, baseada na sorotipagem, multilocus enzimático (MEE) e ribotipagem de N. meningitidis sorogrupo C isoladas de paciente com doença meningocócica no Rio Grande do Sul (RS) e Santa Catarina (SC), onde o Centro de Controle Epidemiológico do Ministério da Saúde detectou um aumento do número de casos de doença meningocócica (DM) devido a este sorogrupo nos últimos 2 anos (1992-1993). Nós demonstramos que a DM devido a cepas de N. meningitidis sorogrupo C no RS e SC que ocorreram nos últimos 4 anos foi devido principalmente por um clone (ET 40), com isolados indistinguíveis por sorogrupo, sorotipo, subtipo e até por ribotipagem. Um pequeno número de casos que não foram devidos a cepas do ET 40 representaram um grupo geneticamente relacionado, que provavelmente é uma nova linhagem gerada do clone ET 40, referido como complexo ET 11 A. Nós também analisamos cepas de N. meningitidis sorogrupo C isoladas na grande São Paulo em 1976 como um grupo representativo do primeiro ano pós-epidêmico na região. A ribotipagem, bem como MEE, puderam fornecer informações sobre as características clonais das cepas isoladas no período pós-epidêmico e também no Sul do Brasil. As cepas de 1976 possuem mais similaridades com as cepas endêmicas atuais do que com as cepas epidêmicas (1992-1993) por ribotipagem, sensibilidade a sulfonamida e MEE. Em conclusão, sorotipagem com anticorpos monoclonais (C:2b:P1.3), MEE (complexo ET11 e ET11A) e ribotipagem usando a enzima de restrição ClaI, foram úteis em caracterizar estas cepas epidêmicas de N. meningitidis relacionadas com o aumento da incidência da DM em diferentes estados do sul do Brasil. É muito provável que estas cepas de N. meningitidis sorogrupo C possuam pouca ou nenhuma correlação genética com as cepas epidêmicas sorogrupo C de 1971-1975. A similaridade genética dos membros do complexo ET 11 e ET 11A foram confirmadas por ribotipagem usando-se 3 enzimas de restrição.

In the present study we report the results of an analysis, based on serotyping, multilocus enzyme electrophoresis (MEE), and ribotyping of N. meningitidis serogroup C strains isolated from patients with meningococcal disease (MD) in Rio Grande do Sul (RS) and Santa Catarina (SC) States, Brazil, as the Center of Epidemiology Control of Ministry of Health detected an increasing of MD cases due to this serogroup in the last two years (1992-1993). We have demonstrated that the MD due to N.meningitidis serogroup C strains in RS and SC States occurring in the last 4 years were caused mainly by one clone of strains (ET 40), with isolates indistinguishable by serogroup, serotype, subtype and even by ribotyping. One small number of cases that were not due to an ET 40 strains, represent closely related clones that probably are new lineages generated from the ET 40 clone referred as ET 11A complex. We have also analyzed N.meningitidis serogroup C strains isolated in the greater São Paulo in 1976 as representative of the first post epidemic year in that region. The ribotyping method, as well as MEE, could provide useful information about the clonal characteristics of those isolates and also of strains isolated in south Brazil. The strains from 1976 have more similarity with the actual endemic than epidemic strains, by the ribotyping, sulfonamide sensitivity, and MEE results. In conclusion, serotyping with monoclonal antibodies (C:2b:P1.3), MEE (ET 11 and ET 11A complex), and ribotyping by using ClaI restriction enzyme (Rb2), were useful to characterize these epidemic strains of N.meningitidis related to the increased incidence of MD in different States of south Brazil. It is mostly probable that these N.meningitidis serogroup C strains have poor or no genetic corelation with 1971-1975 epidemic serogroup C strains. The genetic similarity of members of the ET 11 and ET 11A complex were confirmed by the ribotyping method by using three restriction endonucleases.