Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay

Raquel de A Maschmann, Mirela Verza, Marcia S N Silva, Rosa Dea Sperhacke, Marta O. Ribeiro, Philip Noel Suffys, Harrison Magdinier Gomes, Enrico Tortoli, Fiorella Marcelli, Arnaldo Zaha, Maria Lucia R Rossetti

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)139-145
Number of pages7
JournalMemorias do Instituto Oswaldo Cruz
Volume106
Issue number2
Publication statusPublished - Mar 2011

Keywords

  • Drug resistance
  • Reverse dot blot hybridization
  • Rifampin
  • rpoB
  • Tuberculosis

ASJC Scopus subject areas

  • Microbiology (medical)

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