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Braz. j. infect. dis ; 17(6): 667-671, Nov.-Dec. 2013. ilus, tab
Artículo en Inglés | LILACS | ID: lil-696968

RESUMEN

OBJECTIVE: To evaluate multiplex allele specific polymerase chain reaction as a rapid molecular tool for detecting multidrug-resistant tuberculosis. METHODS: Based on drug susceptibility testing, 103 isolates were multidrug-resistant tuberculosis and 45 isolates were sensitive to isonicotinylhydrazine and rifampin. Primers were designed to target five mutations hotspots that confer resistance to the first-line drugs isoniazid and rifampin, and multiplex allele specific polymerase chain reaction was performed. Whole-genome sequencing confirmed drug resistance mutations identified by multiplex allele specific polymerase chain reaction. RESULTS: DNA sequencing revealed that 68.9% of multidrug-resistant strains have point mutations at codon 315 of the katG gene, 19.8% within the mabA-inhA promoter, and 98.0% at three hotspots within rpoB. Multiplex allele specific polymerase chain reaction detected each of these five mutations, yielding 82.3% sensitivity and 100% specificity for isoniazid resistance, and 97.9% sensitivity and 100% specificity for rifampin resistance as compared to drug susceptibility testing. CONCLUSIONS: The results show that multiplex allele specific polymerase chain reaction is an inexpensive and practical method for rapid detection of multidrug-resistant tuberculosis in developing countries.


Asunto(s)
Humanos , Antituberculosos/farmacología , Reacción en Cadena de la Polimerasa Multiplex , Mycobacterium tuberculosis/genética , Tuberculosis Resistente a Múltiples Medicamentos/diagnóstico , ADN Bacteriano/análisis , Pruebas de Sensibilidad Microbiana , Técnicas de Diagnóstico Molecular , Reacción en Cadena de la Polimerasa Multiplex/economía , Mycobacterium tuberculosis/efectos de los fármacos , Mutación Puntual , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Tuberculosis Resistente a Múltiples Medicamentos/microbiología
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