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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)
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Multidrug-resistant tuberculosis: Rapid molecular detection with MTBDRplus® assay in clinical samples
Tuberculose multirresistente: Detecção directa em amostras respiratórias com o método de genética molecular MTBDRplus®
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Rita Macedo1,
Corresponding author
rita.macedo@csalcantara.min-saude.pt

Correspondence to/Correspondência: Laboratório de Saúde Pública – Micobacteriologia/ Tuberculose, Administração Regional de Saúde de Lisboa e Vale do Tejo, I.P., Rua Pedro Cálmon, n.° 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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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 drugresistant 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 MTBDRplus® assay. The samples from which we isolated resistant strains (36.8%) showed specific mutations associated with the correspondent resistant phenotype.

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

Key-words:
Tuberculosis
Multi-drug resistance
Laboratory diagnosis
Resumo

Uma das principais problematicas no controlo da tuberculose e o aparecimento de casos de tuberculose multirresistente (TB-MR) e tuberculose extensivamente resistente (TB-XDR). A deteccao precoce da resistencia a farmacos, directamente a partir de amostras respiratorias, e essencial para que se assegure o tratamento atempado, adequado e eficaz da tuberculose, bem como para prevenir a disseminacao destes casos de especial gravidade.

O nosso objectivo foi avaliar a sensibilidade e comparar os resultados obtidos com um metodo de genetica molecular disponivel comercialmente – MTBDRplus® – e o isolamento, identificacao e testes de sensibilidade classicos, directamente a partir de amostras respiratorias. Estudamos 68 amostras, com baciloscopia positiva.

O MTBDRplus® permitiu identificar, directamente a partir das amostras respiratorias, o complexo Mycobacterium tuberculosis em todos os casos em que o estudo cultural e a identificacao classica chegaram a esse mesmo resultado. Nas amostras em que culturalmente isolamos estirpes sensiveis, com o MTBDRplus® encontramos sempre perfis geneticos do tipo selvagem (63,2%). Relativamente as amostras em que culturalmente isolamos estirpes resistentes, com o MTBDRplus® encontramos sempre perfis geneticos com mutacoes ou com ausencia do perfil do tipo selvagem (36,8%).

Este estudo permitiu concluir que o MTBDRplus® assegura a deteccao rapida de resistencias a farmacos em estirpes do complexo M. tuberculosis, com resultados totalmente sobreponiveis aos obtidos com os metodos bacteriologicos classicos.

Palavras-chave:
Tuberculose
Multirresistencia
Diagnostico laboratorial.
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