Detection of mutations in Mycobacterium tuberculosis genome determining resistance to fluoroquinolones by hybridization on biological microchips.
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.
Antonova
O V
OV
V. A. Engel'gard Institute of Molecular Biology, Russian Academy of Sciences, Moscow. markova@biochip.ru
Gryadunov
D A
DA
Lapa
S A
SA
Kuz'min
A V
AV
Larionova
E E
EE
Smirnova
T G
TG
Nosova
E Y
EY
Skotnikova
O I
OI
Chernousova
L N
LN
Moroz
A M
AM
Zasedatelev
A S
AS
Mikhailovich
V M
VM
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
D000995:Antitubercular Agents; D024841:Fluoroquinolones