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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 106, No. 2, 2011, pp. 139-145
Bioline Code: oc11023
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 106, No. 2, 2011, pp. 139-145

 en Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay
de A Maschmann, Raquel; Verza, Mirela; Silva, Marcia S.N.; Sperhacke, Rosa Dea; Ribeiro, Marta O.; Suffys, Philip Noel; Gomes, Harrison Magdinier; Tortoli, Enrico; Marcelli, Fiorella; Zaha, Arnaldo & Rossetti, Maria Lucia R.

Abstract

We used a colorimetric reverse dot blot hybridization (CRDH) assay to detect the presence of mutations in a specific region of the rpoB gene, associated with rifampin (RIF) resistance, in a panel of 156 DNAs extracted from 103 RIF-sensitive and 53 RIF-resistant cultures of Mycobacterium tuberculosis. When compared with the antimicrobial susceptibility test (AST), the sensitivity and specificity of the CRDH were 92.3% and 98.1%, respectively. When compared with sequencing, the sensitivity and specificity of the CRDH were 90.6% and 100%, respectively. To evaluate the performance of the assay directly in clinical specimens, 30 samples from tuberculosis patients were used. For these samples, the results of the CRDH were 100% consistent with the results of the AST and sequencing. These results indicate that the rate of concordance of the CRDH is high when compared to conventional methods and sequencing data. The CRDH can be successfully applied when a rapid test is required for the identification of RIF resistance in M. tuberculosis.

Keywords
drug resistance - reverse dot blot hybridization - rifampin - rpoB - tuberculosis

 
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