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au:Telles, Maria Alice da Silva

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Gargalos e recomendações para a incorporação de novas tecnologias na rede pública laboratorial de tuberculose no Brasil
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Telles, Maria Alice da Silva

; Menezes, Alexandre

; Trajman, Anete

Jornal Brasileiro de Pneumologia

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dec 2012, Volume 38 N. 6 Pages 766 - 770

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Novas tecnologias para o diagnóstico da tuberculose foram recentemente recomendadas pela Organização Mundial da Saúde (OMS). Algumas recomendações da OMS incluem a elaboração de um plano estratégico para a adequação da rede, investimentos em supervisão e controle de qualidade, implementação de um sistema de gerenciamento de ambiente laboratorial, sem o que nenhuma nova tecnologia poderá ser eficazmente incorporada. Realizamos um levantamento da rede laboratorial de tuberculose no Brasil para identificar possíveis gargalos para a incorporação dessas tecnologias. Identificamos escassez de recursos para supervisão e controle de qualidade, baixa solicitação de culturas, ausência de sistemas eficazes de informação laboratorial e o desconhecimento da rede periférica municipal quanto às necessidades futuras na infraestrutura.

The World Health Organization (WHO) has recently recommended new technologies for the diagnosis of tuberculosis. The WHO recommendations include the development of a strategic plan for bringing the network up to grade; investment in supervision and quality control; and implementation of a system of laboratory environmental management. Without those measures having been taken, no new technology can be effectively incorporated. We surveyed the tuberculosis laboratory network in Brazil in order to identify possible bottlenecks for the incorporation of new technologies. We identified a lack of resources allocated to supervision and quality control; a low number of requests for cultures; a lack of effective laboratory information systems; and a lack of awareness regarding the future infrastructure needs of the laboratory network at the municipal level.

5391 downloads

Susceptibility of Mycobacterium tuberculosis to first-line antimycobacterial agents in a Brazilian hospital: assessing the utility of the tetrazolium (MTT) microplate assay
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Ferrari, Michela De Luca

; Telles, Maria Alice da Silva

; Ferrazoli, Lucilaine

; Levy, Carlos Emílio

; Villares, Maria Cecília Barison

; Moretti, Maria Luiza

; Resende, Mariângela Ribeiro

Memórias do Instituto Oswaldo Cruz

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aug 2010, Volume 105 N. 5 Pages 661 - 664

Abstract: En | Text: En | PDF: En

We conducted a cross-sectional, hospital-based study between January 2006-March 2008 to estimate the resistance of Mycobacterium tuberculosis to first-line drugs in patients with tuberculosis at a Brazilian hospital. We evaluated the performance of the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) microplate assay compared with the Bactec-MGIT 960™ system for mycobacteria testing. The prevalence of resistance in M. tuberculosis was 6.7%. Multidrug-resistance [resistance to rifampicin (RMP) and isoniazid (INH)], INH-resistance and streptomycin (SM)-resistance accounted for 1%, 3.8% and 3.8% of all resistance, respectively, and all isolates were susceptible to ethambutol (EM). The resistance was primary in four cases and acquired in three cases and previous treatment was associated with resistance (p = 0.0129). Among the 119 M. tuberculosis isolates, complete concordance of the results for INH and EM was observed between the MTT microplate and Bactec-MGIT 960TM methods. The observed agreement for RMP was 99% (sensitivity: 90%) and 95.8% for SM (sensitivity 90.9%), lower than those for other drugs. The MTT colourimetric method is an accurate, simple and low-cost alternative in settings with limited resources.

2219 downloads Cited 1 time in SciELO

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Telles, Maria Alice da Silva

Jornal Brasileiro de Pneumologia

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feb 2010, Volume 36 N. 1 Pages 156 - 156

Abstract: En | Text: En | PDF: En

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Cord factor detection and macroscopic evaluation of mycobacterial colonies: an efficient combined screening test for the presumptive identification of Mycobacterium tuberculosis complex on solid media
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Simeão, Fernanda Cristina dos Santos

; Chimara, Erica

; Oliveira, Rosângela Siqueira

; Yamauchi, Jonas Umeoka

; Latrilha, Fábio Oliveira

; Telles, Maria Alice da Silva

Jornal Brasileiro de Pneumologia

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dec 2009, Volume 35 N. 12 Pages 1212 - 1216

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO: A diferenciação rápida entre Mycobacterium tuberculosis e micobactérias não-tuberculosas é fundamental para os pacientes coinfectados com tuberculose e HIV. Para tanto, utilizamos duas metodologias em nosso laboratório: detecção do fator corda e PCR-restriction enzyme analysis (PRA). O objetivo do estudo foi avaliar a acurácia desse teste de triagem em meio sólido como um método rápido para a identificação presuntiva do complexo M. tuberculosis, considerando custos e tempo de resultado. MÉTODOS: Foram processadas 152 cepas pelo teste de triagem combinado, que consistiu da detecção do fator corda por microscopia (esfregaço corado por Ziehl-Neelsen) e avaliação do aspecto macroscópico das colônias, e PRA (padrão ouro). Os custos foram estimados através da obtenção dos preços dos insumos necessários para a realização de cada teste. RESULTADOS: A acurácia da detecção do fator corda foi de 95,4% (IC95%: 90,7-98,1%) e a do teste de triagem combinado foi de 99,3% (IC95%: 96,4-100%). O custo da detecção do fator corda foi de R$ 0,60 e do PRA de R$ 16,00. Os resultados da detecção do fator corda estão prontos em 2 dias, ao passo que os de PRA necessitam de 4 dias. CONCLUSÕES: A identificação presuntiva de M. tuberculosis usando o aspecto macroscópico das colônias em conjunto com a detecção de fator corda por microscopia é um teste simples, rápido e de baixo custo. Recomendamos o teste de triagem combinado para rapidamente identificar M. tuberculosis em sítios com poucos recursos financeiros e em laboratórios menos equipados, enquanto se aguarda a identificação definitiva por métodos moleculares ou bioquímicos.

OBJECTIVE: The rapid differentiation between Mycobacterium tuberculosis and nontuberculous mycobacteria is fundamental for patients co-infected with tuberculosis and HIV. To that end, we use two methods in our laboratory: detection of cord factor and PCR-restriction enzyme analysis (PRA). The objective of this study was to evaluate the accuracy of a screening test on solid medium as a rapid method for the presumptive identification of M. tuberculosis complex, considering costs and turnover time. METHODS: A total of 152 strains were submitted to a combined screening test, consisting of the detection of cord factor under microscopy (Ziehl-Neelsen staining) and evaluation of the macroscopic aspect of colonies, as well as to PRA, which was used as the gold standard. Costs were estimated by calculating the price of all of the materials needed for each test. RESULTS: The overall accuracy of cord factor detection alone was 95.4% (95% CI: 90.7-98.1%), and that of the combined screening test was 99.3% (95% CI: 96.4-100%). Cord factor detection costs US$ 0.25, whereas the PRA costs US$ 7.00. Results from cord factor detection are ready in 2 days, whereas PRA requires 4 days to yield results. CONCLUSIONS: The presumptive identification of M. tuberculosis using the macroscopic evaluation of colonies combined with cord factor detection under microscopy is a simple, rapid and inexpensive test. We recommend the combined screening test to rapidly identify M. tuberculosis in resource-poor settings and in less well-equipped laboratories while awaiting a definite identification by molecular or biochemical methods.

8228 downloads

III Diretrizes para Tuberculose da Sociedade Brasileira de Pneumologia e Tisiologia
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Conde, Marcus Barreto

; Melo, Fernando Augusto Fiuza de

; Marques, Ana Maria Campos

; Cardoso, Ninarosa Calzavara

; Pinheiro, Valeria Goes Ferreira

; Dalcin, Paulo de Tarso Roth

; Machado Junior, Almério

; Lemos, Antonio Carlos Moreira

; Netto, Antônio Ruffino

; Durovni, Betina

; Sant'Anna, Clemax Couto

; Lima, Dinalva

; Capone, Domenico

; Barreira, Draurio

; Matos, Eliana Dias

; Mello, Fernanda Carvalho de Queiroz

; David, Fernando Cezar

; Marsico, Giovanni

; Afiune, Jorge Barros

; Silva, José Roberto Lapa e

; Jamal, Leda Fátima

; Telles, Maria Alice da Silva

; Hirata, Mário Hiroyuki

; Dalcolmo, Margareth Pretti

; Rabahi, Marcelo Fouad

; Cailleaux-Cesar, Michelle

; Palaci, Moises

; Morrone, Nelson

; Guerra, Renata Leborato

; Dietze, Reynaldo

; Miranda, Silvana Spíndola de

; Cavalcante, Solange Cesar

; Nogueira, Susie Andries

; Nonato, Tatiana Senna Galvão

; Martire, Terezinha

; Galesi, Vera Maria Nader

; Dettoni, Valdério do Valle

Jornal Brasileiro de Pneumologia

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oct 2009, Volume 35 N. 10 Pages 1018 - 1048

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Diariamente novos artigos científicos sobre tuberculose (TB) são publicados em todo mundo. No entanto, é difícil para o profissional sobrecarregado na rotina de trabalho acompanhar a literatura e discernir o que pode e deve ser aplicado na prática diária juntos aos pacientes com TB. A proposta das "III Diretrizes para TB da Sociedade Brasileira de Pneumologia e Tisiologia (SBPT)" é revisar de forma crítica o que existe de mais recente na literatura científica nacional e internacional sobre TB e apresentar aos profissionais da área de saúde as ferramentas mais atuais e úteis para o enfrentamento da TB no nosso país. As atuais "III Diretrizes para TB da SBPT" foram desenvolvidas pela Comissão de TB da SBPT e pelo Grupo de Trabalho para TB a partir do texto das "II Diretrizes para TB da SBPT" (2004). As bases de dados consultadas foram LILACS (SciELO) e PubMed (Medline). Os artigos citados foram avaliados para determinação do nível de evidência científica, e 24 recomendações sobre TB foram avaliadas, discutidas por todo grupo e colocadas em destaque. A primeira versão das "III Diretrizes para TB da SBPT" foi colocada no website da SBPT para consulta pública durante três semanas, e as sugestões, críticas e o nível de evidência da referência científica que as embasavam foram avaliados e discutidos antes de serem incorporadas ou não ao texto final.

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.

112639 downloads Cited 23 times in SciELO

Monitoramento em cabine de segurança biológica: manipulação de cepas e descontaminação em um laboratório de micobactérias
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Ueki, Suely Yoko Mizuka

; Chimara, Erica

; Yamauchi, Jonas Umeoka

; Latrilha, Fábio de Oliveira

; Simeão, Fernanda Cristina dos Santos

; Moniz, Letícia Lisboa

; Giampaglia, Carmen Maria Saraiva

; Telles, Maria Alice da Silva

Jornal Brasileiro de Patologia e Medicina Laboratorial

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aug 2008, Volume 44 N. 4 Pages 263 - 269

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVOS: Verificar evidências da formação de aerossóis durante a manipulação de cepas de micobactérias para teste de sensibilidade às drogas (ANT) e identificação (TIP) e o efeito da descontaminação com solução de álcool 70% e luz ultravioleta (UV) na cabine de segurança biológica (CSB), após os procedimentos laboratoriais. MÉTODOS: Uma placa foi exposta na CSB durante os procedimentos de ANT e TIP. Ao término, a CSB foi limpa e descontaminada com álcool 70% e exposta à luz UV por 15 minutos. Após esse tempo outra placa foi exposta por duas horas, somente com a ventilação da CSB ligada. As placas foram incubadas a 37°C e observadas por 30 dias. Os esfregaços das colônias isoladas foram corados pelas técnicas de Ziehl Neelsen e Gram, e as colônias de bacilo álcool-ácido-resistente (BAAR) foram identificadas pelos métodos tradicionais. RESULTADOS: Nas 38 placas expostas durante o ANT, cresceram micobactérias em 10 placas (26,3%), fungos em uma (2,6%) e outros bacilos em duas (5,3%). Das placas com micobactérias, oito (80%) foram identificadas como M. tuberculosis e duas (20%) tiveram identificação inconclusiva. Mesmo após a descontaminação com álcool 70% e uso de UV, cresceram fungos em duas placas (5,3%) e cocos em outras duas (5,3%). Nas 30 placas colocadas nas CSB durante a TIP, cresceram micobactérias em 10 placas (33,3%), fungos em duas (6,6%), cocos em uma (3,4%) e uma mistura de micobactérias e outro bacilo em uma (3,4%). Não houve crescimento nas placas expostas após descontaminação das CSB com álcool a 70% e uso de UV ao término da TIP. CONCLUSÃO: Durante os procedimentos houve formação de aerossóis contendo micobactérias, fato que ficou comprovado pelo crescimento de colônias de micobactérias nas placas expostas. Técnicas laboratoriais adequadas devem ser respeitadas para minimizar a formação de aerossóis como também observar a experiência dos profissionais, a manutenção de um programa de capacitação e a manutenção periódica da CSB.

OBJECTIVES: To verify the evidence of aerosol formation during the manipulation of mycobacteria strains for susceptibility (ST) and identification tests (IT) as well as the decontamination effect of alcohol solution 70% and ultraviolet (UV) radiation in biological safety cabinets (BSC) after laboratory procedures. METHODS: One plate was exposed in a BSC during ST and IT procedures. Afterwards, the BSC was cleaned and decontaminated with alcohol solution 70% and exposed to UV radiation for 15 minutes. After that, another plate was exposed for two hours, only with the BSC ventilation on. Both plates were incubated at 37°C and observed for 30 days. The smears from the isolated colonies were stained with Ziehl Neelsen and Gram techniques, and acid fast bacilli (AFB) were identified by conventional methods. RESULTS: In 38 plates exposed during ST, there was mycobacteria growth in 10 plates (26.3%), fungi in one (2.6%) and bacilli in two (5.3%). Among those plates that presented mycobacteria growth, eight (80%) were identified as M. tuberculosis and two (20%) had inconclusive identification. Even after decontamination with alcohol solution 70% and UV radiation, two plates presented fungi growth (5.3%) and other two presented cocci growth (5.3%). Among 30 plates exposed during IT procedures, there was mycobacteria growth in 10 of them (33.3%), fungi in two (6.6%), cocci in one (3.4%) and one (3.4%) mixed mycobacteria and another bacillus. No growth was observed when alcohol solution 70% and UV radiation were used for decontamination after IT procedures. CONCLUSION: During the procedures there was aerosol formation with mycobacteria, which was proved by mycobacteria growth on the exposed plates. Not only should adequate laboratory techniques be respected to minimize aerosol formation, but professional expertise, the continuity of capacity programs and periodic BSC maintenance should also be observed.

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Identification of non-tuberculous mycobacteria from the Central Public Health Laboratory from Mato Grosso do Sul and analysis of clinical relevance
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Moraes, Paulo Ricardo de Souza

; Chimara, Erica

; Telles, Maria Alice da Silva

; Ueki, Suely Yoko Misuka

; Cunha, Eunice Atsuko Totumi

; Honer, Michael Robin

; Leão, Sylvia Cardoso

Brazilian Journal of Microbiology

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jun 2008, Volume 39 N. 2 Pages 268 - 272

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Micobactérias não-tuberculosas isoladas no Laboratório Central de Saúde Pública de Mato Grosso do Sul em 2003 e 2004 foram identificadas usando métodos fenotípicos convencionais (TI) e PCR-Restriction Enzyme Analysis (PRA) tendo o gene hsp65 como alvo (PRA-hsp65). Em 15 dos 32 isolados analisados os resultados obtidos com ambos métodos foram concordantes, sendo 8 Mycobacterium avium, 3 M. fortutium, 1 M. kansasii, 1 M. flavescens, 1 M. peregrinum e 1 Nocardia brasiliensis. TI de 12 isolados não foi conclusiva. Perfis não descritos de PRA-hsp65 foram observados com 11 isolados. Dados dos prontuários médicos foram avaliados para inferir a relevância clínica dos isolados.

Non-tuberculous mycobacteria isolated at the Central Public Health Laboratory from Mato Grosso do Sul in 2003 and 2004 were identified by conventional phenotypic methods (TI) and by PCR-Restriction Enzyme Analysis (PRA) using the hsp65 gene as target (PRA-hsp65). With 15 of the 32 analysed isolates, results of both methods were concordant, being 8 Mycobacterium avium, 3 M. fortutium, 1 M. kansasii, 1 M. flavescens, 1 M. peregrinum and 1 Nocardia brasiliensis. TI of 12 isolates was inconclusive. Novel PRA-hsp65 patterns were observed with 11 isolates. Medical data were evaluated for inference of clinical relevance of these isolates.

3149 downloads Cited 1 time in SciELO

Micobactérias não-tuberculosas: diversidade das espécies no estado de São Paulo
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Ueki, Suely Yoko Mizuka

; Martins, Maria Conceição

; Telles, Maria Alice da Silva

; Virgilio, Melissa Curcio

; Giampaglia, Carmen Maria Saraiva

; Chimara, Erica

; Ferrazoli, Lucilaine

Jornal Brasileiro de Patologia e Medicina Laboratorial

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feb 2005, Volume 41 N. 1 Pages 1 - 8

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O gênero Mycobacterium é constituído por espécies do complexo M. tuberculosis e outras denominadas micobactérias não-tuberculosas (MNT). Até o momento, mais de cem MNT foram descritas. Os objetivos deste estudo foram avaliar a diversidade das espécies de MNT identificadas no estado de São Paulo, no período de 1991 a 1997, que antecedeu a expansão da terapia anti-retroviral, e determinar a freqüência dos casos que atenderam alguns critérios bacteriológicos para o diagnóstico das infecções causadas pelas MNT. MATERIAL E MÉTODOS: Foram analisadas 1.892 cepas isoladas de sítios estéreis e não-estéreis de 1.248 pacientes atendidos no estado de São Paulo. RESULTADOS: Do total de pacientes, 1.199 (96,1%) tiveram suas cepas identificadas e 3,9% apresentaram resultados não-conclusivos. As dez espécies encontradas foram o complexo M. avium (MAC), M. kansasii, M. chelonae, M. fortuitum, M. szulgai, M. xenopi, M. marinum, M. gordonae, M. terrae e M. nonchromogenicum. Quarenta e sete (7,8%) casos pulmonares tiveram diagnóstico confirmado pelo isolamento da mesma espécie em três ou mais amostras e 67 (34%) pacientes tiveram o diagnóstico bacteriológico confirmado por isolamento em sítios estéreis. CONCLUSÕES: As espécies de MNT mais freqüentemente isoladas no estado de São Paulo foram MAC e M. kansasii. Uma publicação nacional com recomendações para diagnóstico e tratamento dessas infecções seria fundamental para a conduta correta no diagnóstico e no tratamento de micobacterioses.

INTRODUCTION: Mycobacterium genus includes species of M. tuberculosis complex and others called non-tuberculous mycobacteria (NTM). So far, more than 100 species of NTM have been described. The objectives of this study were to estimate the NTM species diversity among strains isolated in São Paulo State from 1991 to 1997, before the expansion of antiretroviral therapy, and to evaluate the rate of cases that fall under diagnostic criteria for bacteriological diagnosis of NTM infections. MATERIAL AND METHODS: 1,892 strains were isolated from sterile and non-sterile sites of 1,248 patients. RESULTS: 1,199 (96%) were identified as M. avium complex (MAC), M. kansasii, M. chelonae, M. fortuitum, M. szulgai, M. xenopi, M. marinum, M. gordonae, M. terrae or M. nonchromogenicum whereas 49 (3.9%) couldn't be identified up to species. Forty-seven (7,8%) cases from pulmonary sites were confirmed based on strains isolated from three or more samples from the same patient and 67 (34%) patients had NTM isolated from sterile sites, which confirmed the bacteriological diagnosis. CONCLUSIONS: MAC and M. kansasii were the most frequently especies isolated in São Paulo state. National recomendations for the diagnosis and treatment of NTM infections would be useful for correct management of NTM infections.

7002 downloads

Molecular characterization of Mycobacterium kansasii isolates in the State of São Paulo between 1995-1998
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Chimara, Erica

; Giampaglia, Carmen Maria Saraiva

; Martins, Maria Conceição

; Telles, Maria Alice da Silva

; Ueki, Suely Yoko Mizuka

; Ferrazoli, Lucilaine

Memórias do Instituto Oswaldo Cruz

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nov 2004, Volume 99 N. 7 Pages 739 - 743

Abstract: En | Text: En | PDF: En

Mycobacterium kansasii is the most common cause of pulmonary nontuberculous mycobacteria infection and classical identification of this pathogen needs a time consuming phenotypic tests. Polymerase chain reaction-restriction fragment lenght polymorphism analysis (PRA) of the gene enconding for the 65kDa heat shock (hsp65) protein offers an easy, rapid, and inexpensive procedure to identify and subtype M. kansasii isolates. In the present study, we performed a retrospective analysis of patients who had mycobacteria identified on the basis of phenotypic tests by means of a review of database at Mycobacteria Laboratory of the Instituto Adolfo Lutz in the period 1995-1998. A total of 9381 clinical isolates were analyzed of which 7777 (82.9%) were identified as M. tuberculosis complex and 1604 (17.1%) as nontuberculous mycobacteria. Of the 296 M. kansasii isolates, 189 (63.8%) isolates obtained from 119 patients were viable and were analyzed by PRA-hsp65. Hundred eight two (98.9%) were classified as M. kansasii type I. Two isolates were classified as type II and III and five isolates were characterized as other Mycobacterium species. Clinical isolates of M. kansasii in the state of São Paulo was almost exclusively subtype I regardless of HIV status.

2144 downloads Cited 3 times in SciELO

10.

Distribución de patrones PRA en aislamientos clínicos del complejo Mycobacterium avium procedentes de España y Suramérica
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Murcia, Martha Isabel

; Cardoso Leao, Sylvia

; Ritacco, Viviana

; Palenque, Elia

; Siqueira de Oliveira, Rosangela

; Reniero, Ana

; Menendez, Maria Carmen

; da Silva Telles, María Alice

; Hadad, David Jamil

; Barrera, Lucía

; García, María Jesús

Biomédica

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jun 2004, Volume 24 Suppl. 1 Pages 60 - 64

Abstract: Es En | Text: Es En | PDF: Es | PDF: En

La infección por el complejo Mycobacterium avium (MAC) es la infección sistémica más frecuente en la fase terminal del SIDA. Las sondas de ADN disponibles en el mercado para la identificación de micobacterias son muy precisas pero extremadamente costosas. Por eso, la mayoría de los laboratorios clínicos de Latinoamérica aún tipifican micobacterias mediante pruebas fenotípicas que son lentas, laboriosas y poco precisas. En este trabajo se aplicó el análisis del polimorfismo de los fragmentos de restricción del gen hsp65 (PRA) a la identificación de MAC en 163 aislamientos clínicos procedentes de España y Suramérica. El genotipo PRA predominante en cada país fue: M. avium tipo I en Argentina (23/42, 55%) y Brasil (48/72, 67%), M. avium tipo II en España (18/26, 69%) y M. avium tipo III en Colombia (10/23, 43%). Este último genotipo, que aún no fue descrito fuera del continente americano, resultó muy infrecuente en los otros tres países del estudio. Se discuten ventajas e inconvenientes de la aplicación del PRA al diagnóstico micobacteriológico.

Distribution of PRA patterns of clinical isolates of the Mycobacterium avium complex from Spain and South America Mycobacterium avium complex (MAC) infections are the most frequent systemic infections associated with advanced AIDS. DNA probes for accurate identification of mycobacteria are available but are very expensive in many Latin American settings. Consequently, most Latin American diagnostic laboratories employ inaccurate and outdated tests for mycobacteria identification. Therefore, PCR restriction analysis (PRA) of the hsp65 gene was evaluated for the identification of 163 MAC human isolates originated from Spain and South America. The predominant PRA type in each country was: M. avium type I in Argentina (23/42, 55%) and Brazil (48/72, 67%), M. avium type II in Spain (18/26, 69%) and M. avium type III in Colombia (10/ 23, 43%). The Colombia frequency is noteworthy, since the PRA type III was quite infrequent in the other three countries. Furthermore, its presence has not been reported outside the Americas. The advantages and disadvantages of PRA in diagnostic mycobacteriology are discussed

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11.

Cord formation and colony morphology for the presumptive identification of Mycobacterium tuberculosis complex
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Monteiro, Paulo Henrique Tasso

; Martins, Maria Conceição

; Ueki, Suely Yoko Mizuka

; Giampaglia, Carmen Maria Saraiva

; Telles, Maria Alice da Silva

Brazilian Journal of Microbiology

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jun 2003, Volume 34 N. 2 Pages 171 - 174

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O resultado da identificação convencional do complexo Mycobacterium tuberculosis (MT) é demorado. O objetivo deste estudo foi avaliar um teste de triagem para identificação presuntiva de MT e agilização da informação preliminar do resultado, baseado na análise da morfologia da colônia e da visualização de corda no exame microscópico do esfregaço feito da cultura. De janeiro de 1998 a dezembro de 1999 foram analisadas 3056 culturas no Instituto Adolfo Lutz, São Paulo, Brasil. As culturas classificadas como micobactérias não tuberculosas foram submetidas ao teste de identificação por método convencional (testes de crescimento em cultura e bioquímicos) e as MT foram submetidas ao teste de sensibilidade às drogas. Os resultados de sensibilidade, especificidade, valores preditivos positivo e negativo, obtidos com a comparação da identificação presuntiva de MT com o teste de triagem proposto e a identificação final com o método convencional, foram respectivamente 98,9, 86,9, 97,8 e 93,0%. Concluiu-se que a análise da morfologia da colônia e da presença de corda no exame microscópico possibilitou a identificação presuntiva de MT. Este teste pode ser usado pelo laboratório para informação preliminar do resultado presuntivo e para direcionar de maneira prática e com baixo custo, os testes adicionais de sensibilidade às drogas ou de identificação de espécie que serão necessários.

The identification of Mycobacterium tuberculosis complex (MT), using non-molecular methods, is time-consuming. The objective of this study was to evaluate a screening test for the presumptive identification of MT, which could potentially decrease laboratory turn-around time for reporting preliminary results. From January 1998 to December 1999, 3056 cultures were analysed at the Mycobacterial Laboratory, Instituto Adolfo Lutz, São Paulo, Brasil. The screening test consisted of observation of colony morphology on Löwenstein Jensen medium and evaluation of cord formation on smear microscopy from those positive cultures. After the screening test, the cultures identified as non-tuberculous mycobacteria were identified to species by conventional methods (growth on culture and biochemical tests). Those identified as MT were submitted to drug susceptibility tests. The presumptive identification of MT using the proposed screening test, when compared with conventional tests, presented 98.9, 86.9, 97.8 and 93.0% of sensitivity, specificity, positive and negative predictive values, respectively. The conclusion is that it is possible to make a presumptive identification of MT using visual analysis of colony morphology and cord formation on microscopy examination. This method could be used to report the presumptive identification of MT and to guide laboratory decisions regarding susceptibility and identification tests with little cost and in a very practical way.

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12.

Standartization of broth microdilution method for Mycobacterium tuberculosis
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Leite, Clarice Queico Fujimura

; Beretta, Ana Laura Remédio Zeni

; Anno, Ivone Shizuko

; Telles, Maria Alice da Silva

Memórias do Instituto Oswaldo Cruz

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jan 2000, Volume 95 N. 1 Pages 127 - 129

Abstract: En | Text: En | PDF: En

Indirect drug susceptibility tests of Mycobacterium tuberculosis was done to investigate the accuracy and feasibility of a broth microdilution method (BMM) for determining minimal inhibitory concentrations of conventional drugs against M. tuberculosis. Test drugs included isoniazid (H), rifampicin (R), ethambutol (E), streptomycin (S) and pyrazinamide (Z). Fifty isolates of M. tuberculosis from patients who had never received drug therapy, and H37Rv strain for control, were evaluated in the system. When comparing this method with the gold standard proportional method in Lowenstein-Jensen medium, sensitivity of 100% for all drugs and specifities of 91, 100, 96, 98 and 85% were observed respectively for H, R, E, S and Z. The BMM was read faster (14-20 days) than the proportional method (20-28 days). The microdilution method evaluated allows the testing of multiple drugs in multiple concentrations. It is easy to perform and does not require special equipment or expensive supplies. In contrast to radiometric method it does not use radioactive material.

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13.

Mycobacterium avium complex (MAC) isolated from AIDS patients and the criteria required for its implication in disease
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Hadad, David Jamil

; Palhares, Maria Cecília de Almeida

; Placco, Anna Luiza Nunes

; Domingues, Carmem Silvia Bruniera

; Castelo Filho, Adauto

; Ferrazoli, Lucilaine

; Ueki, Sueli Yoko Mizuka

; Telles, Maria Alice da Silva

; Martins, Maria Conceição

; Palaci, Moisés

Revista do Instituto de Medicina Tropical de São Paulo

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oct 1995, Volume 37 N. 5 Pages 375 - 383

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Anterior a pandemia de AIDS, o Complexo Mycobacterium avium (MAC) era responsável pela maioria das vezes, por pneumopatias acometendo pacientes com doença pulmonar crônica de base como enfisema e bronquite crônica36. Em 1981, com o advento da síndrome de imunodeficiência adquirida (SIDA), o MAC passou a representar uma das doenças bacterianas mais frequentes em pacientes com esta síndrome, sendo a doença disseminada a principal forma de manifestação clínica da infecção8. Entre Janeiro de 1989 e Fevereiro de 1991, no Setor de Micobactérias do Instituto Adolfo Lutz em São Paulo, o MAC foi isolado de 103 pacientes a partir do cultivo de diferentes espécimes estéreis e não estéreis processados, coletados de 2.304 pacientes atendidos no Centro de Referência e Treinamento AIDS e/ou Instituto de Infectologia Emilio Ribas. A doença disseminada foi diagnosticada em 29 destes, com base no isolamento do MAC a partir do sangue e/ou aspirado de medula óssea. Os outros 74 pacientes foram agrupados nas categorias altamente (5), moderadamente (26) e pouco sugestiva de doença (43) de acordo com os postulados de DAVIDSON (1989)10. Os diferentes critérios para valorizar o seu isolamento de espécimes estéreis e não estéreis são discutidos.

Before the AIDS pandemia, the Mycobacterium avium complex (MAC) was responsible in most cases for the pneumopathies that attack patients with basic chronic pulmonary diseases such as emphysema and chronic bronchitis36. In 1981, with the advent of the acquired immunodeficiency syndrome (AIDS), MAC started to represent one of the most frequent bacterial diseases among AIDS patients, with the disseminated form of the disease being the major clinical manifestation of the infection8. Between January 1989 and February 1991, the Section of Mycobacteria of the Adolfo Lutz Institute, São Paulo, isolated MAC from 103 patients by culturing different sterile and no-sterile processed specimens collected from 2304 patients seen at the AIDS Reference and Training Center and/or Emilio Ribas Infectology Institute. Disseminated disease was diagnosed in 29 of those patients on the basis of MAC isolation from blood and/or bone marrow aspirate. The other 74 patients were divided into categories highly (5), moderately (26) and little suggestive of disease (43) according to the criteria of DAVIDSON (1989)10. The various criteria for MAC isolation from sterile and non-sterile specimens are discussed.

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14.

Bacterial agents isolated from cerebrospinal fluid of patients with Acquired Immunodeficiency Syndrome (AIDS) and neurological complications
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Landgraf, Ilka Maria

; Palaci, Moisés

; Vieira, Maria de Fátima Paiva

; Ueki, Sueli Yoko Mizuka

; Martins, Maria Conceição

; Sato, Daisy Nakamura

; Ferrazoli, Lucilaine

; Telles, Maria Alice Silva

; Gorla, Maria Cecília Outeiro

; Vaz, Tania Mara Ibelli

; Lorenço, Rosemarie

; Marques, Lilian Regina Macelloni

Revista do Instituto de Medicina Tropical de São Paulo

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dec 1994, Volume 36 N. 6 Pages 491 - 496

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Amostras de líquido cefalorraquidiano (LCR) de 2083 pacientes com Síndrome de Imunodeficiência Adquirida (SIDA) e complicações neurológicas foram examinados durante um período de 7 anos (1984-1990). A porcentagem de pacientes que tiveram pelo menos um agente bacteriano cultivado do LCR foi de 6,2%. Mycobacterium tuberculosis foi o mais frequentemente isolado (4,3%), seguido do complexo Mycobacterium avium ou MAC (0,7%), de Pseudomonas spp (0,5%), Enterobacter spp (0,4%), e Staphylococcus aureus (0,3%). Entre 130 pacientes com cultura positiva, de 89 (68,5%) foi isolado M. tuberculosis e de 15 (11,6%) MAC. A frequência de isolamentos bacterianos aumentou de 1988 (5,2%) a 1990 (7,2%), particularmente devido ao maior isolamento de MAC. Os agentes bacterianos foram mais frequentemente isolados de pacientes na faixa etária de 21-30 anos e de mulheres (p<0.05).

Cerebrospinal fluid (CSF) samples from 2083 patients with acquired immunodeficiency syndrome (AIDS) and neurological complications were bacteriologically examined during a period of 7 years (1984-1990). The percentage of patients who had at least one bacterial agent cultured from the CSF was 6.2%. Mycobacterium tuberculosis was the most frequently isolated agent (4.3%), followed by Mycobacterium avium complex or MAC (0.7%), Pseudomonas spp (0.5%), Enterobacter spp (0.4%), and Staphylococcus aureus (0.3%). Among 130 culture positive patients, 89 (68.5%) had M. tuberculosis and 15 (11.6%) had MAC. The frequency of bacterial isolations increased from 1988 (5.2%) to 1990 (7.2%), partly due to the increase in MAC isolations. Bacterial agents were more frequently isolated from patients in the age group 21-30 years and from women (p<0.05).

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