title: Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay.
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.
authors.affiliation: Fundação Estadual de Produção e Pesquisa em Saúde, Porto Alegre, RS, Brasil. raquel368@gmail.com
authors.forename: Raquel de A
authors.initials: Rde A
authors.lastname: Maschmann
authors.forename: Mirela
authors.initials: M
authors.lastname: Verza
authors.forename: Marcia Sn
authors.initials: MS
authors.lastname: Silva
authors.forename: Rosa Dea
authors.initials: RD
authors.lastname: Sperhacke
authors.forename: Marta O
authors.initials: MO
authors.lastname: Ribeiro
authors.forename: Philip Noel
authors.initials: PN
authors.lastname: Suffys
authors.forename: Harrison Magdinier
authors.initials: HM
authors.lastname: Gomes
authors.forename: Enrico
authors.initials: E
authors.lastname: Tortoli
authors.forename: Fiorella
authors.initials: F
authors.lastname: Marcelli
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Zaha
authors.forename: Maria Lucia R
authors.initials: ML
authors.lastname: Rossetti
mesh_terms: D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology*; D012680:Sensitivity and Specificity
chemical_list: D000904:Antibiotics, Antitubercular; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C505805:rpoB protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin