Silvana Spindola de Miranda

III Diretrizes para Tuberculose da Sociedade Brasileira de Pneumologia e Tisiologia

New scientific articles about tuberculosis (TB) are published daily worldwide. However, it is difficult for health care workers, overloaded with work, to stay abreast of the latest research findings and to discern which information can and should be used in their daily practice on assisting TB patients. The purpose of the III Brazilian Thoracic Association (BTA) Guidelines on TB is to critically review the most recent national and international scientific information on TB, presenting an updated text with the most current and useful tools against TB to health care workers in our country. The III BTA Guidelines on TB have been developed by the BTA Committee on TB and the TB Work Group, based on the text of the II BTA Guidelines on TB (2004). We reviewed the following databases: LILACS (SciELO) and PubMed (Medline). The level of evidence of the cited articles was determined, and 24 recommendations on TB have been evaluated, discussed by all of the members of the BTA Committee on TB and of the TB Work Group, and highlighted. The first version of the present Guidelines was posted on the BTA website and was available for public consultation for three weeks. Comments and critiques were evaluated. The level of scientific evidence of each reference was evaluated before its acceptance for use in the final text.

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.

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.

Perfil funcional de pacientes portadores de seqüela de tuberculose de um hospital universitário

OBJECTIVE: To describe data related to the pulmonary function of patients with sequelae of pulmonary tuberculosis, pleural tuberculosis or both. METHODS: In the outpatient clinic of a university hospital, 218 patients were evaluated. Of those 218, 56 had sequelae of tuberculosis (pulmonary, pleural or both), and 162 had other types of tuberculosis. All patients were evaluated in the pulmonary function laboratory between February 2000 and July 2004, and 43 were found to be eligible for inclusion in the study. Patients with a history of asthma, chronic pulmonary obstructive disease, cardiac insufficiency, collagen diseases, silicosis or thoracic surgery, as well as those for whom spirometry yielded unacceptable results or was not performed, were excluded. The lung fields were divided into six zones, and radiographic results were classified by degree: I (involvement of only one zone with no cavitation); II (involvement of two or three zones or of one zone with cavitation); or III (extensive involvement of three or more zones with or without cavitation). RESULTS: The final study sample comprised 50 patients, 44 (88%) of whom had pulmonary tuberculosis. The most prevalent form (17/50; 34%) was mixed ventilatory disturbance. Severe disturbances were more significant in degree III radiographs (p = 0.0002) and normal pulmonary function was predominant among patients presenting degree I and II radiographs (p = 0.002). CONCLUSION: The early discovery and treatment of tuberculosis contribute to reduce the number of cases, as well as the incidence of tuberculosis sequelae, thereby improving the quality of life of tuberculosis patients. Further studies, involving longitudinal, sequential analysis and larger samples of patients with tuberculosis sequelae, should be conducted in referral centers in Brazil.

Characteristics of tuberculosis in the state of Minas Gerais, Brazil: 2002-2009

OBJECTIVE: To analyze the profile of tuberculosis cases reported between 2002 and 2009 in the state of Minas Gerais, Brazil, according to sociodemographic, clinical, and laboratory characteristics, as well as to comorbidities and mortality. METHODS: This was a descriptive, epidemiological study based on data obtained from the Brazilian Case Registry Database and the Brazilian Mortality Database for the 2002-2009 period. RESULTS: There were 47,285 reported cases of tuberculosis, corresponding to a mean incidence of 22.3/100,000 population. The individuals diagnosed with tuberculosis were predominantly in the 20- to 49-year age bracket and male (62.4% and 67.0%, respectively). Individuals with a low level of education accounted for 18.5% of the cases. New cases, cases of recurrence, and cases of retreatment accounted for 83.7%, 5.7%, 5.7%, respectively. The rates of cure and treatment noncompliance were 66.2% and 11.2%, respectively; multidrug-resistant tuberculosis was identified in 0.2% of the cases; and the mortality rate was 12.9%. The directly observed treatment, short-course (DOTS) strategy was applied in 21.8% of the cases. Sputum smear microscopy and culture were performed in only 73.9% and 12.9% of the cases, respectively. Chest X-rays were performed in 90.5% of the cases. Pulmonary tuberculosis was the predominant form (in 83.9%). Comorbidity with alcoholism, HIV infection, and diabetes mellitus were identified in 17.2%, 8.3%, and 3.8%, respectively. CONCLUSIONS: During the study period, the numbers of new cases, cases of treatment noncompliance, and deaths were high, comorbidities were common, and there was a failure to perform adequately basic tests for the diagnosis of tuberculosis. Multidisciplinary approaches, expanded use of the DOTS strategy, better knowledge of the distribution of tuberculosis, and improvements in the databases are needed in order to achieve better control of the disease in the state of Minas Gerais.

Spoligotyping of clinical Mycobacterium tuberculosis isolates from the state of Minas Gerais, Brazil

We performed spoligotyping on 114 strains of the Mycobacterium tuberculosis (Mtb) complex that had been isolated from patients in Minas Gerais Health Units during 2004. A total of 82/114 (72%) clinical isolates were clustered and 32/114 (28%) were unique. Seven shared types containing nine strains were newly created. A total of nine patterns corresponded to unreported orphan strains, as evaluated against all of the strains recorded in the SITVIT2 proprietary database in the Institut Pasteur de la Guadeloupe. The major clades were composed of isolates that belong to the following genotypes: Latin-America and Mediterranean (63/114, 55.3%) (the ill-defined T superfamily) (12/114, 10.5%), Haarlem (8/114, 7%), X clade (6/114, 5.3%), S clade (3/114, 2.6%) and the East-African Indian and Manu types, each with 1/114 (0.9%) isolates. A considerable number of strains (n = 20, 17.5%) showed patterns that did not fall within any of the previously described major clades. We conclude the bulk of tuberculosis (TB) (92/114, 80.7%) in our location is recent evolutionary strains that belong to the principal genetic groups 2/3. Further studies on epidemiology of TB are required to understand Mtb biodiversity and TB transmission in this region.

Evaluation of six different DNA extraction methods for detection of Mycobacterium tuberculosis by means of PCR-IS6110: preliminary study

BackgroundDevelopments in molecular detection and strain differentiation of members of Mycobacterium tuberculosis complex have proved to be useful. The DNA extraction method influences the amplification efficiency, causing interference on the sensitivity and respective inhibitors. The aim of this study was to standardize a simple and fast DNA extraction method, providing DNA amplification by IS6110-PCR effectively free from undue interferences.FindingsThe efficiency of the six different protocols tested in M. tuberculosis cultures has varied from 75% to 92.5%. This preliminary study evaluating the IS6110 PCR sensitivity and specificity was developed in DNA extracted from microscope slides, and achieved 100% of efficiency.ConclusionsDNA extraction by Chelex + NP-40 method from both, cultures of M. tuberculosis and smear slides, resulted in good quantity of interference free DNA, especially in samples with low concentrations of genetic material; therefore, such technique may be used for the molecular diagnosis of tuberculosis.

Diagnóstico de resistência do Mycobacterium tuberculosis à rifampicina utilizando-se da reação em cadeia da polimerase

A resistencia do Mycobacterium tuberculosis aos tuberculostaticos tem surgido como grande ameaca a Saude Publica. A resistencia a rifampicina pode ser considerada como um marcador para a multi-resistencia a farmacos e tem sido atribuida a mudancas estruturais da RNA polimerase, produto de expressao do gene rpoB. Os pacientes portadores dessas cepas tem baixa perspectiva frente ao tratamento. Os testes convencionais de sensibilidade aos farmacos realizados em cultura do Mtb requerem varias semanas para o crescimento. Por este motivo, a Reacao em Cadeia da Polimerase (PCR), metodo de baixo custo e que pode reduzir o tempo para o diagnostico, representa alternativa viavel e promissora. Neste artigo estao descritos os metodos mais comumente empregados na deteccao de mutantes resistentes a rifampicina baseados na PCR, como analise de Polimorfismo Conformacional de Fita Simples (Single-Strand Conformation Polymorphism, SSCP), PCR Heteroduplex e INNO-LIPA. Recentemente, padronizou-se a tecnica de PCR em baixa estringencia, usando um unico iniciador (Low Stringency Single Specific Primer, LSSP), que se mostrou um metodo rapido e sensivel na deteccao de mutacoes no gene rpoB.

Low-Stringency Single-Specific-Primer PCR as a Tool for Detection of Mutations in the rpoB Gene of Rifampin-Resistant Mycobacterium tuberculosis

ABSTRACT By the low-stringency single-specific-primer PCR technique, a highly sensitive and rapid method for diagnosis of rifampin resistance in Mycobacterium tuberculosis was established. Seven rifampin-resistant and five rifampin-susceptible specimens were analyzed. Rifampin resistance was determined by MIC measurement. A complex electrophoretic pattern consisting of many bands was obtained for both susceptible and rifampin-resistant isolates. The same pattern was obtained for all of the susceptible specimens, but differences between resistant and susceptible isolates were found. DNA sequencing showed that a particular mutation produces a specific electrophoretic pattern.

Positive tuberculin test and risk of infection by Mycobacterium tuberculosis in a tuberculosis clinic settled in an upright building, in Minas Gerais, Brazil

Antecedentes: La tuberculosis ocupacional causa ausentismo laboral. Conocer la reactividad tuberculinica de los trabajadores de la salud ayuda a mejorar las medidas de control para evitar la infeccion laboral con M. tuberculosis. Objetivo: Estudiar la reactividad tuberculinica de un grupo de trabajadores de la salud de un centro de referencia de tuberculosis que esta ubicado en una edificacion vertical. Material y Metodos: Se realizo una prueba de tuberculina a 251 empleados de una clinica de tuberculosis, que no tenian antecedentes o sospecha de tuberculosis y quienes no habian sido sometidos previamente a la prueba. Aquellos sujetos que no respondieron a tuberculina fueron sometidos a una nueva prueba para evaluar la reaccion de refuerzo. Resultados: El 51% de los sujetos estudiados tuvieron una reaccion tuber-culinica positiva. El fenomeno de refuerzo se observo en 69 de 190 sujetos (35,7%). La tasa de seroconversion en aquellos sujetos con una tuberculina negativa en que se repitio la prueba la cabo de un ano, fue de 5,1%. Conclusiones: En este grupo de trabajadores de la salud expuestos a M. tuberculosis, hay una alta tasa de reactividad negativa a tuberculina.

Brazilian Response to Global End TB Strategy : The National Tuberculosis Research Agenda.

Afranio Kritski[1],[2], Draurio Barreira[3], Ana Paula Junqueira-Kipnis[1],[4], Milton Ozorio Moraes[5], Maria Martha Campos[1],[6], Wim Mauritz Degrave[7], Silvana Spindola Miranda[1],[8], Marco Aurelio Krieger[1],[9], Erica Chimara[1],[10], Carlos Morel[11], Margareth Pretti Dalcolmo[1],[12], Ethel Leonor Noia Maciel[1],[13], Maria do Socorro Nantua Evangelista[3],[14], Teresa Scatena Villa[1],[15], Mauro Sanchez[1],[16], Fernanda Dockhorn Costa[3], Inacio Queiroz[17], Martha Maria Oliveira[1],[11], Ruy Souza Junior[3], Jose Roberto Lapa e Silva[1],[2] and Antonio Ruffi no-Netto[1],[18]