title: [Detection of wild type isolates and point mutations in genome of isoniazid-resistant Mycobacteria tuberculosis].
abstract: The isoniazid resistance of mycobacteria tuberculosis (MBT) is associated with point mutations in the codon 315 of katG gene of MBT. The two PCR-techniques for detection of point mutations in codon 315 have been developed. A use of two sets of primers comprising the additional competitive blocking primer with 3'-terminal phosphate group (in order to eliminate unspecific amplification) allowed to identify most frequent point mutations AGC-->ACC and AGC-->AGA in the codon 315. PCR with a set of two primers one of which contained 5 LNA-monomers allowed to discriminate between wild type and isoniazid-resistant MBT isolates; any of 6 known mutations in codon 315 of kafG gene being detected. A structure and purity of the LNA-containing oligonucleotides (length of 17 nucleotides) was characterized by MALDI-TOF mass spectrometry; the duplex formed by two LNA-containing complementary oligonucleotides (length of 17 b.p.) was anlyzed by thermal denaturation.
authors.forename: O Iu
authors.initials: OIu
authors.lastname: Limanskaia
authors.forename: T N
authors.initials: TN
authors.lastname: Mukhina
authors.forename: V N
authors.initials: VN
authors.lastname: Stepanshina
authors.forename: I G
authors.initials: IG
authors.lastname: Shemiakin
authors.forename: X
authors.initials: X
authors.lastname: Wu
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: T V
authors.initials: TV
authors.lastname: Fecenko
authors.forename: V A
authors.initials: VA
authors.lastname: Pokrovskiĭ
authors.forename: A P
authors.initials: AP
authors.lastname: Limanskiĭ
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D002374:Catalase / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D016680:Genome, Bacterial; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D017354:Point Mutation
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; D007538:Isoniazid