title: Detection of mutations in Mycobacterium tuberculosis genome determining resistance to fluoroquinolones by hybridization on biological microchips.
abstract: We developed a method of identification of Mycobacterium tuberculosis with simultaneous evaluation of the sensitivity to fluoroquinolones on a biological microchip array. The method of multiplex two-staged PCR followed by hybridization of a biochip makes it possible to detect 8 mutant variants of gyrA gene occurring in fluoroquinolone-resistant strains (approximately 85% all resistant forms) within 1 day. Using this method we analyzed 107 cultures isolated from patients with tuberculosis and 78 sputum samples. Mutations in gyrA gene were detected in 48 (92%) resistant strains. Natural S95T polymorphism in gyrA gene was detected in all resistant and in 76% sensitive strains. The sensitivity and specificity of the proposed method calculated on the basis of the analysis of sputum samples (n=78) were 94 and 100%, respectively.
authors.affiliation: V. A. Engel'gard Institute of Molecular Biology, Russian Academy of Sciences, Moscow. markova@biochip.ru
authors.forename: O V
authors.initials: OV
authors.lastname: Antonova
authors.forename: D A
authors.initials: DA
authors.lastname: Gryadunov
authors.forename: S A
authors.initials: SA
authors.lastname: Lapa
authors.forename: A V
authors.initials: AV
authors.lastname: Kuz'min
authors.forename: E E
authors.initials: EE
authors.lastname: Larionova
authors.forename: T G
authors.initials: TG
authors.lastname: Smirnova
authors.forename: E Y
authors.initials: EY
authors.lastname: Nosova
authors.forename: O I
authors.initials: OI
authors.lastname: Skotnikova
authors.forename: L N
authors.initials: LN
authors.lastname: Chernousova
authors.forename: A M
authors.initials: AM
authors.lastname: Moroz
authors.forename: A S
authors.initials: AS
authors.lastname: Zasedatelev
authors.forename: V M
authors.initials: VM
authors.lastname: Mikhailovich
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001483:Base Sequence; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D024841:Fluoroquinolones / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D006824:Hybridization, Genetic; D046228:Microarray Analysis / Q000379:methods*; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics; D018088:Tuberculosis, Multidrug-Resistant / Q000188:drug therapy / Q000235:genetics / Q000382:microbiology
chemical_list: D000995:Antitubercular Agents; D024841:Fluoroquinolones