@article {Alvarez FigueroaP513, author = {Maria Alvarez Figueroa and German Shipulin}, title = {Application of the novel technique next gene sequencing for the determination of mycobacterium tuberculosis resistance in clinical samples}, volume = {42}, number = {Suppl 57}, elocation-id = {P513}, year = {2013}, publisher = {European Respiratory Society}, abstract = {Genetic investigation of drug-resistant tuberculosis (DR-TB) is extremely difficult due to involvement of a large number of genes, the presence of compensatory and unknown mutations, the lack of epidemiological studies in different geographical areas and migration. The more careful determination of Mycobacterium tuberculosis (MTB) sensitivity is required in order to make the informed choice of drugs. Existing molecular methods possess restricted capacity due to their ability to determine only a limited number of mutations or drugs.We try to apply a novel technique Next Gene Sequencing (NGS) to identify the sequence of rpoB as a model of MTB separate genes. This approach allows to perform simultaneous analysis of all known target genes of drugs used for TB treatment in up to 95 clinical samples.We randomly selected from the strain bank of Central Research Institute for Epidemiology 12 sputum samples and 9 clinical isolates which had been cultivated from these samples. All samples had been previously investigated by cultural method and Sanger sequencing.NGS results were scrutinized and revealed that 2 samples had a wild type, 4 - a single mutation (2 - 531L, 1 - 526N, 1 - 516V), 3 - multiple mutations (526N+533P, (516Y+531L)+wild, (511R+516Y+531L) +wild). The study demonstrates a complete coincidence between the NGS results and earlier obtained data. The interesting fact is that ratio between different mutations in samples from third group retained in clinical samples and isolates.Our study demonstrates that NGS provides a perspective tool for the future management of DR-TB with much more efficiency than any of the previously used methods.}, issn = {0903-1936}, URL = {https://erj.ersjournals.com/content/42/Suppl_57/P513}, eprint = {https://erj.ersjournals.com/content/42/Suppl_57/P513.full.pdf}, journal = {European Respiratory Journal} }