Skrypnyk A. Molecular-genetic differentiation of mycobacteria isolated in Ukraine, and their phylogenetic interrelationships

The object of the research - husbandry animal tuberculosis: tuberculosis infection agents and atypical mycobacteria. The goal - to identify the species structure of mycobacteria isolated in different regions of Ukraine, to study their cultural-morphological properties, phylogenetic interrelationships and to develop the system of their molecular-genetic identification and species differentiation. Methods of the research - bacteriological, molecular-biological, genetic, statistical. Theoretical and practical results: the comparative evaluation of diagnostic effectiveness of molecular-genetic methods for Mycobacteria species differentiation was carried out, the percentage of atypical species and subspecies isolated from cattle was determined, phylogenetic tree incorporated 17 Mycobacteria species was built. Species of 125 Mycobacteria strains was determined, our investigations revealed the circulation on the territory of Ukraine such species: M. caprae, M. frederіksbergensе, M. engbackіі, M. dorіcum, M.parascrofulaceum, M. hassіacum, M. confluentіs, M. elephantіs. The typing M. аvіum strains to the subspecies level of M. avіum subsp. homіnіssuіs и M. avіum subsp. avіum was conducted. The method for mycobacterial DNA extraction from the bacterial mass for use in PCR was developed. The system of Mycobacteria species differentiation and their genetic relatedness detection by genotyping was worked out. Genus-specific kit for Mycobacteria detection by PCR was developed. Novelty: The comparative evaluation of diagnostic effectiveness of molecular-genetic methods for Mycobacteria species differentiation and molecular epizootiology was carried out. Genus-specific kit for Mycobacteria detection by PCR, the method of mycobacterial DNA extraction, the method for diagnostics of animal tuberculosis and mycobacteriosis. Degree of implementation: Normative documentation for the kit, registration certificate (№2742-14-03-00-07, 06.10.2007) and methodical recommendations are approved by The State Department of Veterinary Medicine of Ukraine. Field of application - veterinary medicine.