title: Detection of ethambutol-resistant Mycobacterium tuberculosis strains by multiplex allele-specific PCR assay targeting embB306 mutations.
abstract: We describe a multiplex allele-specific (MAS)-PCR assay to detect simultaneously mutations in the first and third bases of the embB gene codon 306ATG. These mutations are known to confer ethambutol (EMB) resistance in the majority of clinical Mycobacterium tuberculosis isolates worldwide. The mutated bases are revealed depending on the presence or absence of the respective indicative fragments amplified from the embB306 wild-type allele. Initially optimized on purified DNA samples, the assay was tested on crude cell lysates and auramine-stained sputum slide DNA preparations with the same reproducibility and interpretability of the generated profiles in agarose gel electrophoresis. Since EMB resistance is generally linked to multiple-drug resistance (MDR), the MAS-PCR assay for EMB resistance detection can be used in clinical laboratory practice in areas with a high prevalence and a high transmission rate of MDR-EMB-resistant tuberculosis.
authors: Laboratory of Molecular Microbiology, Pasteur Institute, 14, Mira Street, St. Petersburg 197101, Russia. miv@IM4520.spb.edu
authors: Igor
authors: I
authors: Mokrousov
authors: Olga
authors: O
authors: Narvskaya
authors: Elena
authors: E
authors: Limeschenko
authors: Tatiana
authors: T
authors: Otten
authors: Boris
authors: B
authors: Vyshnevskiy
mesh_terms: D000483:Alleles; D000995:Antitubercular Agents / Q000494:pharmacology*; D001483:Base Sequence; D017931:DNA Primers; D004352:Drug Resistance, Microbial; D004977:Ethambutol / Q000494:pharmacology*; D006801:Humans; D016296:Mutagenesis; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D010430:Pentosyltransferases / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping
chemical_list: D000995:Antitubercular Agents; D017931:DNA Primers; D004977:Ethambutol; D010430:Pentosyltransferases; C101967:arabinosyltransferase