2021  1,540
2020  1,374
2019  1,023
2018  0,932
2017  0,977
2016  0,799
2015  0,662
2014  0,740
2013  0,739
2012  0,637
2011  0,658
2010  0,654
2009  0,570
2008  0,849
2007  0,805
2006  0,330
2005  0,435
2004  0,623
2003  0,567
2002  0,641
2001  0,490
2000  0,477
1999  0,762
1998  0,785
1997  0,507
1996  0,518
1995  0,502
Vol 48(2014) N 2 p. 214-226; DOI 10.1134/S0026893314020186 Full Text

D.V. Zimenkov1*, E.V. Kulagina1, O.V. Antonova1, S.A. Surzhikov1, Yu.A. Bespyatykh2, E.A. Shitikov2, E.N. Ilina2, V.M. Mikhailovich1, A.S. Zasedatelev1,3, D.A. Gryadunov1

Analysis of the Genetic Determinants of Multidrug and Extensive Drug Resistance in Mycobacterium tuberculosis with the Use of an Oligonucleotide Microchip

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia
2Institute of Physico-Chemical Medicine, Federal Medical-Biological Agency, Moscow, 119435, Russia
3Moscow Physico-Technical Institute, Dolgoprudnyi, Moscow Region, 141700, Russia

Received - 2013-07-12; Accepted - 2013-09-04

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 multidrug-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.

Mycobacterium tuberculosis, multidrug and extensive drug resistance, multiplex PCR, hybridization, oligonucleotide microchip