Detection of second-line drug resistance in Mycobacterium tuberculosis using oligonucleotide microarrays

BMC Infectious Diseases

Dmitry A Gryadunov¹  Vladimir M Mikhailovich¹  Alexander S Zasedatelev¹  Sergey A Popov³  Ludmila Y Krylova²  Yulia D Isaeva²  Alexey V Kuz’min³  Olga V Antonova¹  Danila V Zimenkov¹ 

[1] Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia;Moscow Scientific and Clinical Antituberculosis Center, Department of Health of Moscow, Moscow, Russia;Research Institute for Phthisiopulmonology, I. M. Sechenov Moscow Medical Academy, Moscow, Russia

关键词: XDR-TB;    Oligonucleotide microarrays;    Extensively drug-resistant tuberculosis;    Hybridisation;    Biochips;   

Others  :  1148196
DOI  :  10.1186/1471-2334-13-240

 received in 2012-12-24, accepted in 2013-05-21,  发布年份 2013

【 摘 要 】

Background

The steady rise in the spread of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) requires rapid and reliable methods to identify resistant strains. The current molecular methods to detect MTB resistance to second-line drugs either do not cover an extended spectrum of mutations to be identified or are not easily implemented in clinical laboratories. A rapid molecular technique for the detection of resistance to second-line drugs in M. tuberculosis has been developed using hybridisation analysis on microarrays.

Methods

The method allows the identification of mutations within the gyrA and gyrB genes responsible for fluoroquinolones resistance and mutations within the rrs gene and the eis promoter region associated with the resistance to injectable aminoglycosides and a cyclic peptide, capreomycin. The method was tested on 65 M. tuberculosis clinical isolates with different resistance spectra that were characterised by their resistance to ofloxacin, levofloxacin, moxifloxacin, kanamycin and capreomycin. Also, a total of 61 clinical specimens of various origin (e.g., sputum, bronchioalveolar lavage) were tested.

Results

The sensitivity and specificity of the method in the detection of resistance to fluoroquinolones were 98% and 100%, respectively, 97% and 94% for kanamycin, and 100% and 94% for capreomycin. The analytical sensitivity of the method was approximately 300 genome copies per assay. The diagnostic sensitivity of the assay ranging from 67% to 100%, depending on the smear grade, and the method is preferable for analysis of smear-positive specimens.

Conclusions

The combined use of the developed microarray test and the previously described microarray-based test for the detection of rifampin and isoniazid resistance allows the simultaneous identification of the causative agents of MDR and XDR and the detection of their resistance profiles in a single day.

【 授权许可】

   
2013 Zimenkov et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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