Drug susceptibility testing of Mycobacterium tuberculosis using next generation sequencing and Mykrobe software

Introduction. Mycobacterium tuberculosis is the causative agent of tuberculosis. Drug susceptibility testing performed by phenotypic and molecular tests. Commonly used for drug automated BACTEC system in a liquid culture medium. line probe tests was introduced almost 15 years ago. Recently whole genome sequencing (WGS) analysis M. strains demonstrated that genotyping drug-resistance could be accurately performed. Several software tools were developed.Our study aimed to perform whole-genome on phenotypically confirmed multi-drug resistant (MDR) strains, identify drug-resistant mutations compare profiles with assay results.Materials methods. We 34 MDR Bulgarian strains. Phenotypic system. For first- second-line tuberculostatics, we applied Geno Type MTBDR plus v.1.0 и sl v.1.0. Sequencing MiSeq. Generated FASTQ files analyzed known drugresistant platform Mykrobe v.0.8.1.Results. All three methods — using system, genetic applying test gave comparable sensitivity/resistance results studied proven rifampicin isoniazid-resistant 100% software. The C-15T mutation marker isoniazid resistance SIT41 spoligotype. observed 75% (21/28) agreement between ethambutol resistance. Phenotypically, 87% (n = 27) are streptomycin, but only 59% 19) Comparing genotypic ofloxacin, amikacin kanamycin, coincidence results.Conclusions. Whole-genome approach relatively expensive laborious useful detailed such as epidemiological testing.