16S-ITS-23S rRNA operon segment sequencing provides necessary and sufficient conditions for bacterial species-specific identification

Introduction. Sequencing of the 16S rRNA gene is predominant method for assessing microbial communities and strain molecular identification. The short reads (2nd generation sequencing)-based technology does not allow analysis beyond gene. taxonomic verification level samples usually remains at genus or even family level. Currently, there have been proposed latest versions long-read technologies (Oxford Nanopore MinION, PacBio) amplicon sequencing near-complete ribosomal operon, including genes 16S, 23S, 5S, internal transcribed spacer (ITS). At moment, this approach has sufficiently studied, in addition, it involves PCR amplification a very extended DNA region (more than 4000 bp-long). Materials methods. collection non-tuberculous mycobacteria strains their primary identification was carried out years 20192021. were obtained by inoculation positive cultures from Bactec MGIT 960 bacteriological analyzer lacking MPT64 antigen TB Identification Test (Becton Dickinson, USA) on Lowenstein-Jensen medium. Preliminary species with Speed-oligo Mycobacteria kit (Vircell, Spain) according to manufacturers protocol. In work, both known newly developed universal bacterial primers flanking gene, ITS, beginning 23S are used. present study, used flank start Results discussion. Sanger amplicons (about 2000 bp) shows sufficient determine up 8 isolated humans that caused clinically bacteriologically confirmed diseases. operon fragment containing most considered us as an approbation methodological study material high degree degradation (necrotic foci, etc.). results indicate significantly higher resolution classical sequencing.