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dc.creatorRuiz Pérez, Maitees
dc.creatorTorres Sánchez, María Josées
dc.creatorLlanos Rodríguez, Ana Cristinaes
dc.creatorArroyos, Aurelioes
dc.creatorPalomares Folía, José Carloses
dc.creatorAznar Martín, Javieres
dc.identifier.citationRuíz Pérez, M., Torres Sánchez, M.J., Llanos Rodríguez, A.C., Arroyos, A., Palomares Folia, J.C. y Aznar Martín, J. (2004). Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR. Journal of Clinical Microbiology, 42 (4), 1585-1589.
dc.identifier.issn0095-1137 (impreso)es
dc.identifier.issn1098-660X (electronico)es
dc.description.abstractOur objective was to evaluate the feasibility of a molecular assay based on a real-time PCR technique, carried out with a LightCycler instrument (Roche Biochemicals), to identify Mycobacterium tuberculosis bacilli and to detect rifampin and isoniazid resistance in DNA extracts from sputum samples. We studied three genes: rpoB, which is associated with rifampin resistance, and katG and inhA, which are associated with isoniazid resistance. A total of 205 sputum samples collected from 108 patients diagnosed with pulmonary tuberculosis with positive auramine-rhodamine-staining (AR) sputum samples, were tested. The sensitivities of the LightCycler PCR assay for the positive AR specimens was 97.5% (200 of 205) for rpoB and inhA genes and 96.5% (198 of 205) for the katG gene. For the total number of patients tested, the sensitivity was 100% (108 of 108 patients) for rifampin, whereas the sensitivity was 98.1% (106 of 108 patients) for isoniazid. Full agreement was found with the Bactec MGIT 960 method and the genotype inferred from the LightCycler data for rifampin. The phenotypic method for isoniazid reported 13 resistant strains (≥0.1 μg/ml). In seven (53.8%) strains there was a concordance between both methods, but we found that six (46.2%) strains reported as resistant by the phenotypic method were determined to be susceptible by real-time PCR. For the 75 strains reported as susceptible by the phenotypic method, the concordance with the LightCycler data was 100%. Our results demonstrate that rifampin-resistant M. tuberculosis could be detected in DNA extracted from auramine-rhodamine-positive sputum samples in a single-tube assay that took less than 3 h to perform for a collection of auramine-rhodamine-positive specimens obtained from patients with culture-documented pulmonary tuberculosis. Similarly, this occurs in half of the isoniazid-resistant M. tuberculosis DNA extracted from auramine-rhodamine-positive
dc.description.sponsorshipFondo de Investigación Sanitaria, Ministerio de Sanidad y Consumo FIS 01/051es
dc.publisherAmerican Society for Microbiologyes
dc.relation.ispartofJournal of Clinical Microbiology, 42 (4), 1585-1589.
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internacional*
dc.titleDirect Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCRes
dc.contributor.affiliationUniversidad de Sevilla. Departamento de Microbiologíaes
dc.relation.projectIDFIS 01/051es
dc.journaltitleJournal of Clinical Microbiologyes
dc.contributor.funderMinisterio de Sanidad y Consumo. España

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