Brazilian Journal of Pulmonology

ISSN (on-line): 1806-3756 | ISSN (printed): 1806-3713

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Evaluation of a genetic probe (Gen-Probe Accuprobe® system) in comparison to traditional methods for identifying members of the Mycobacterium tuberculosis complex

Avaliação de uma sonda genética (Sistema Accuprobe, Gen Probe®) para identificação de organismos do complexo Mycobacterium tuberculosis, em comparação com métodos tradicionais de caracterização

Delurce Tadeu de Araujo Spada, Manoel Armando Azevedo dos Santos, Elisabete A. Almeida, Marcos Augusto, Maria Idemar Pedrosa Albarral, Fernando Augusto Fiuza de Melo

J Bras Pneumol.2005;31(3):219-224

Abstract PDF PT PDF EN Portuguese Text

Background: The appearance of tuberculosis/human immunodeficiency virus co-infection and the growing number of diseases caused by nontuberculous mycobacteria, as well as the confusion that these can cause in relation to emerging multidrug-resistant strains, require more accurate and rapid laboratory results, not only in the isolation of strains but also in their identification. Objective: A comparative study evaluating a new tool of molecular identification, which uses a genetic probe based on the 16S rDNA sequence of the Mycobacterium tuberculosis gene (Gen-Probe Accuprobe® Gen Probe, Inc.), and the classic methodology. Method: Fifty-five Mycobacterium strains, isolated from the sputum of patients treated at a tuberculosis reference clinic, were selected for study. Subcultures were performed in three tubes: one submitted to genetic identification, one analyzed through classical tests (production and accumulation of niacin; growth in the Lowenstein-Jensen medium with the inhibitor agents p-nitrobenzoic acid and thiophene-2-carboxylic acid hydrazide added), and one held in reserve. Results: The probe identified 51 cases as belonging to the M. tuberculosis complex (one associated with M. kansasii) and the other 4 as nontuberculous mycobacteria, later identified as M. kansasii (3) and M. avium (1). Using traditional methods, 47 samples were identified as belonging to the M. tuberculosis complex, 4 were classified as fitting the profile of nontuberculous mycobacteria (in agreement with the genetic probe results), and 4 were unidentified, 1 of which presented the exact characteristics that 2 mycobacterium species have in common. Conclusion: The benefits of the molecular biology technique justify its implementation and routine use, in combination with classical methods, in a high-traffic clinic where complex cases of tuberculosis are treated.

 


Keywords: Mycobacterium tuberculosis. Molecular probe techniques.

 


Sputum culture yield: comparing an automated diagnostic system to Löwenstein-Jensen medium in the diagnosis of pulmonary tuberculosis

Rendimento da cultura de escarro na comparação de um sistema de diagnóstico automatizado com o meio de Lowenstein-Jensen para o diagnóstico da tuberculose pulmonar

Elisabete Aparecida de Almeida, Manoel Armando Azevedo dos Santos, Jorge Barros Afiune, Delurce Tadeu de Araújo Spada, Fernando Augusto Fiuza de Melo

J Bras Pneumol.2005;31(3):231-236

Abstract PDF PT PDF EN Portuguese Text

Background: Tuberculosis continues to be a global health problem. Objective: To evaluate an automated system designed to diagnose tuberculosis, comparing it to sputum microscopy and culture in Löwenstein-Jensen medium. Method: A comparative study using 844 sputum samples, collected between September and December of 1999 at a reference center for tuberculosis in São Paulo, Brazil, to draw distinctions between the results obtained through the use of the automated system and those obtained through sputum microscopy and culture in Löwenstein-Jensen medium. Results: Of the 844 samples evaluated, 27.1% tested positive for acid-fast bacilli, and 72.9% tested negative. In Löwenstein-Jensen culture, 34.7% were positive and 63% were negative, compared with 37.1% positivity and 56.9% negativity using the automated system. Sensitivity was 98.1% for the automated system and 91.9% for Löwenstein-Jensen culture. Specificity and positive predictive value were 100% for both methods. Negative predictive value was 98.9% for the automated system and 95.5% for Löwenstein-Jensen culture. The degree of accuracy was 99.3% for the automated system and 97% for Löwenstein-Jensen culture, and the Kappa was 0.99 for the automated system and 0.94 for Löwenstein-Jensen culture. The difference between the mean time to detection of mycobacteria using the automated system (10.5 days) and that found using Löwenstein-Jensen culture (34.7 days) was statistically significant. Conclusion: The difference between the culture yield obtained using the automated system and that achieved with Löwenstein-Jensen culture was statistically significant. Mean time to detection of mycobacteria was significantly shorter with the automated system. The higher yield provided by this new system justifies its use in a reference center for tuberculosis in São Paulo

 


Keywords: Mycobacterium tuberculosis. Diagnostic. Automated systems.

 


 

 


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CNPq, Capes, Ministério da Educação, Ministério da Ciência e Tecnologia, Governo Federal, Brasil, País Rico é País sem Pobreza
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SCS Quadra 01, Bloco K, Salas 203/204 Ed. Denasa. CEP: 70.398-900 - Brasília - DF
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E-mails: jbp@jbp.org.br
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