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In house colorimetric reverse hybridisation assay for detection of the mutation most frequently associated with resistance to isoniazid in Mycobacterium tuberculosis

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Author(s): Mirela Verza | Raquel de Abreu Maschmann | Márcia Susana Nunes Silva | Elis Regina Dalla Costa | Marta Osório Ribeiro | Franciele Rosso | Philip Noel Suffys | Enrico Tortoli | Fiorella Marcelli | Arnaldo Zaha | Maria Lucia Rosa Rossetti

Journal: Memórias do Instituto Oswaldo Cruz.
ISSN 0074-0276

Volume: 104;
Issue: 5;
Start page: 710;
Date: 2009;
Original page

Keywords: drug resistance | isoniazid | katG315 | reverse hybridization assay | 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.
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