TY - JOUR
T1 - Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay
AU - Maschmann, Raquel de A
AU - Verza, Mirela
AU - Silva, Marcia S N
AU - Sperhacke, Rosa Dea
AU - Ribeiro, Marta O.
AU - Suffys, Philip Noel
AU - Gomes, Harrison Magdinier
AU - Tortoli, Enrico
AU - Marcelli, Fiorella
AU - Zaha, Arnaldo
AU - Rossetti, Maria Lucia R
PY - 2011/3
Y1 - 2011/3
N2 - 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.
AB - 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.
KW - Drug resistance
KW - Reverse dot blot hybridization
KW - Rifampin
KW - rpoB
KW - Tuberculosis
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M3 - Article
C2 - 21537671
AN - SCOPUS:79955388589
VL - 106
SP - 139
EP - 145
JO - Memorias do Instituto Oswaldo Cruz
JF - Memorias do Instituto Oswaldo Cruz
SN - 0074-0276
IS - 2
ER -