title: In house colorimetric reverse hybridisation assay for detection of the mutation most frequently associated with resistance to isoniazid in Mycobacterium tuberculosis.
abstract: Mutations in the katG gene have been identified and correlated with isoniazid (INH) resistance in Mycobacterium tuberculosis isolates. The mutation AGC-->ACC (Ser-->Thr) at katG315 has been reported to be the most frequent and is associated with transmission and multidrug resistance. Rapid detection of this mutation could therefore improve the choice of an adequate anti-tuberculosis regimen, the epidemiological monitoring of INH resistance and, possibly, the tracking of transmission of resistant strains. An in house reverse hybridisation assay was designed in our laboratory and evaluated with 180 isolates of M. tuberculosis. It could successfully characterise the katG315 mutation in 100% of the samples as compared to DNA sequencing. The test is efficient and is a promising alternative for the rapid identification of INH resistance in regions with a high prevalence of katG315 mutants.
authors.affiliation: Centro de Desenvolvimento Científico e Tecnológico, Fundação Estadual de Produção e Pesquisa em Saúde, Porto Alegre, RS, Brazil.
authors.forename: Mirela
authors.initials: M
authors.lastname: Verza
authors.forename: Raquel de Abreu
authors.initials: Rde A
authors.lastname: Maschmann
authors.forename: Márcia Susana Nunes
authors.initials: MS
authors.lastname: Silva
authors.forename: Elis Regina
authors.initials: ER
authors.lastname: Dalla Costa
authors.forename: Marta Osório
authors.initials: MO
authors.lastname: Ribeiro
authors.forename: Franciele
authors.initials: F
authors.lastname: Rosso
authors.forename: Philip Noel
authors.initials: PN
authors.lastname: Suffys
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 Rosa
authors.initials: ML
authors.lastname: Rossetti
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D002374:Catalase / Q000235:genetics*; D003124:Colorimetry / Q000379:methods; D004269:DNA, Bacterial / Q000032:analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D007538:Isoniazid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009154:Mutation / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; D007538:Isoniazid