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Vol. 15. Issue 3.
Pages 353-365 (May - June 2009)
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Vol. 15. Issue 3.
Pages 353-365 (May - June 2009)
Artigo Original/Original Article
Open Access
Tuberculose multirresistente: Detecção directa em amostras respiratórias com o método de genética molecular MTBDRplus®
Multidrug-resistant tuberculosis: Rapid molecular detection with MTBDRplus® assay in clinical samples
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Rita Macedo1,
Corresponding author
rita.macedo@csalcantara.min-saude.pt

Correspondência / Correspondence to: Laboratório de Saúde Pública – Micobacteriologia/ Tuberculose, Administração Regional de Saúde de Lisboa e Vale do Tejo, I.P., Rua Pedro Cálmon, n.º 25, 1300-455 Lisboa Tel: +351 213602520 Fax: +351 213602525
, António Amorim1, Edna Pereira1
1 Laboratório de Saúde Pública: Micobacteriologia/ Tuberculose, Administração Regional de Saúde de Lisboa e Vale do Tejo, IP/Public Health Laboratory: Mycobacteriology/Tuberculosis, Regional Health Administration, Lisbon and Tagus Valley Health Authority
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Resumo

Uma das principais problemáticas no controlo da tuberculose é o aparecimento de casos de tuberculose multirresistente (TB-MR) e tuberculose extensiva-mente resistente (TB-XDR). A detecção precoce da resistência a fármacos, directamente a partir de amostras respiratórias, é essencial para que se assegure o tratamento atempado, adequado e eficaz da tuberculose, bem como para prevenir a disseminação destes casos de especial gravidade.

O nosso objectivo foi avaliar a sensibilidade e comparar os resultados obtidos com um método de genética molecular disponível comercialmente – MTBDRplus® – e o isolamento, identificação e testes de sensibilidade clássicos, directamente a partir de amostras respiratórias. Estudamos 68 amostras, com baciloscopia positiva.

O MTBDRplus® permitiu identificar, directamente a partir das amostras respiratórias, o complexo Mycobacte-rium tuberculosis em todos os casos em que o estudo cultural e a identificação clássica chegaram a esse mesmo resultado. Nas amostras em que culturalmente iso-lámos estirpes sensíveis, com o MTBDRplus® encontrá-mos sempre perfis genéticos do tipo selvagem (63,2%). Relativamente às amostras em que culturalmente isolá-mos estirpes resistentes, com o MTBDRplus® encontrámos sempre perfis genéticos com mutações ou com au-sência do perfil do tipo selvagem (36,8%).

Este estudo permitiu concluir que o MTBDRplus® assegura a detecção rápida de resistências a fármacos em estirpes do complexo M. tuberculosis, com resultados totalmente sobreponíveis aos obtidos com os mé-todos bacteriológicos clássicos.

Rev Port Pneumol 2009; XV (3): 353-366

Palavras-chave:
Tuberculose
multirresistência
diagnóstico laboratorial
Abstract

Nowadays, the greatest concern of tuberculosis control programmes is the appearance of multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis. Rapid determination of drug resistance in clinical samples, with Mycobacterium tuberculosis complex (MTC), is the prerequisite for initiating effective chemotherapy, ensuring successful treatment of the patient and preventing further spread of drug-resistant isolates.

The aim of our study was to determine the sensitivity of the new MTBDRplus® assay in comparison to culture, identification and classic DST, directly from smear-positive clinical specimens.

A total of 68 smear-positive sputum specimens were processed by both the classical mycobacteriological methods and the molecular assay, MTBDRplus®.

MTBDRplus® assay allowed an accurate identification of MTC species by detection of the specific band in all samples, from which we also isolated and identified MTC strains by culture methods. In the samples from which we isolated susceptible strains (63.2%), wild type patterns were found using MTB-DRplus® assay. The samples from which we isolated resistant strains (36.8%) showed specific mutations associated with the correspondent resistant pheno-type.

Our study indicated that this assay allows rapid detection of resistance, always in agreement with classic methods.

Rev Port Pneumol 2009; XV (3): 353-366

Key-words:
Tuberculosis
multi-drug resistance
laboratory diagnosis
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