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Detection of mutations in Mycobacterium tuberculosis genome determining resistance to fluoroquinolones by hybridization on biological microchips

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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 (∼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.

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Authors and Affiliations

  1. V. A. Engel’gard Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

    O. V. Antonova, D. A. Gryadunov, S. A. Lapa, A. V. Kuz’min, E. E. Larionova, T. G. Smirnova, E. Yu. Nosova, O. I. Skotnikova, L. N. Chernousova, A. M. Moroz, A. S. Zasedatelev & V. M. Mikhailovich

  2. Central Institute of Tuberculosis, Russian Academy of Medical Sciences, Moscow, Russia

    O. V. Antonova, D. A. Gryadunov, S. A. Lapa, A. V. Kuz’min, E. E. Larionova, T. G. Smirnova, E. Yu. Nosova, O. I. Skotnikova, L. N. Chernousova, A. M. Moroz, A. S. Zasedatelev & V. M. Mikhailovich

  3. Moscow Research-and-Practical Center for Tuberculosis Control, Moscow, Russia

    O. V. Antonova, D. A. Gryadunov, S. A. Lapa, A. V. Kuz’min, E. E. Larionova, T. G. Smirnova, E. Yu. Nosova, O. I. Skotnikova, L. N. Chernousova, A. M. Moroz, A. S. Zasedatelev & V. M. Mikhailovich

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  1. O. V. Antonova
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  2. D. A. Gryadunov
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  3. S. A. Lapa
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  4. A. V. Kuz’min
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  5. E. E. Larionova
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  6. T. G. Smirnova
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  7. E. Yu. Nosova
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  8. O. I. Skotnikova
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  9. L. N. Chernousova
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  10. A. M. Moroz
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  11. A. S. Zasedatelev
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  12. V. M. Mikhailovich
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Corresponding author

Correspondence to O. V. Antonova.

Additional information

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Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 1, pp. 115–120, January, 2008

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Antonova, O.V., Gryadunov, D.A., Lapa, S.A. et al. Detection of mutations in Mycobacterium tuberculosis genome determining resistance to fluoroquinolones by hybridization on biological microchips. Bull Exp Biol Med 145, 108–113 (2008). https://doi.org/10.1007/s10517-008-0034-5

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  • DOI: https://doi.org/10.1007/s10517-008-0034-5

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