title: [Analysis of genetic determinants of multidrug and extensively drug-resistant Mycobacterium tuberculosis using oligonucleotide microchip].
abstract: Steadily growing resistance of the tuberculosis causative agent towards a broad spectrum of anti-tuberculosis drugs calls for rapid and reliable methods for identifying the genetic determinants responsible for this resistance. In this study, we present a biochip-based method for simultaneous identification of mutations within rpoB gene associated with rifampin resistance, mutations in katG, inhA, ahpC genes responsible for isoniazid resistance, mutations within the regions of gyrA and gyrB genes leading to fluoroquinolones resistance, and mutations in the rrs gene and the eis promoter region associated with the resistance to kanamycin, capreomycin and amikacin. The oligonucleotide microchip, as the core element of this assay, provides simultaneous identification of 99 mutations in the format "one sample--one PCR--one microchip", and it makes it possible to complete analysis of multi-drug-resistant and extensively drug-resistant tuberculosis within a single day. The tests on 63 Mycobacterium tuberculosis clinical isolates with different resistance profiles using the developed approach allows us to reveal the spectrum of drug-resistance associated mutations, and to estimate the significance of the inclusion of extra genetic loci in the determination of M. tuberculosis drug resistance.
authors: D V
authors: DV
authors: Zimenkov
authors: E V
authors: EV
authors: Kulagina
authors: O V
authors: OV
authors: Antonova
authors: S A
authors: SA
authors: Surzhikov
authors: Iu A
authors: IuA
authors: Bespiatykh
authors: E A
authors: EA
authors: Shitikov
authors: E N
authors: EN
authors: Il'ina
authors: V M
authors: VM
authors: Mikhaĭlovich
authors: A S
authors: AS
authors: Zasedatelev
authors: D A
authors: DA
authors: Griadunov
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015373:Bacterial Typing Techniques; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D027081:DNA Gyrase / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D054908:Extensively Drug-Resistant Tuberculosis / Q000382:microbiology; D024841:Fluoroquinolones / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D057166:High-Throughput Screening Assays / Q000295:instrumentation*; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D007612:Kanamycin / Q000494:pharmacology; D046208:Microchip Analytical Procedures; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D054464:Peroxiredoxins / Q000235:genetics / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D024841:Fluoroquinolones; C505805:rpoB protein, Mycobacterium tuberculosis; D007612:Kanamycin; D010088:Oxidoreductases; D054464:Peroxiredoxins; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; C085257:InhA protein, Mycobacterium; D012321:DNA-Directed RNA Polymerases; D027081:DNA Gyrase; D007538:Isoniazid; D012293:Rifampin