title: Improved real-time PCR for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis clinical isolates.
abstract: In this study we designed two pairs of probes for the detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis with real-time PCR procedures. One pair of probes spans the region between codon 510 and 528 of the rpoB gene, and the other one screens for mutation at the regulatory region of the inhA gene. We have evaluated these probes in combination with two other pairs of probes previously described to detect mutations in 20 susceptible and 53 unique resistant M. tuberculosis clinical isolates. We were able to detect nine different mutations affecting five codons of the rpoB gene, two different mutations at codon 315 of the katG gene and a nucleotide substitution (C209T) in the regulatory region of the inhA gene within two hours turnaround.
authors.affiliation: Departamento de Microbiología, Unidad de Microbiología Molecular, Facultad de Medicina, Sevilla, Spain. mjtorres@us.es <mjtorres@us.es>
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Torres
authors.forename: Antonio
authors.initials: A
authors.lastname: Criado
authors.forename: Maite
authors.initials: M
authors.lastname: Ruiz
authors.forename: Ana C
authors.initials: AC
authors.lastname: Llanos
authors.forename: Jose C
authors.initials: JC
authors.lastname: Palomares
authors.forename: Javier
authors.initials: J
authors.lastname: Aznar
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis*; D024901:Drug Resistance, Multiple, Bacterial; D005260:Female; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D008297:Male; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis / Q000188:drug therapy
chemical_list: D004269:DNA, Bacterial; D007538:Isoniazid; D012293:Rifampin