Chastii T. Microbiological characteristics of fungi of the genus Malassezia and improved methods of laboratorial diagnostics of malasseziosis

Object of research: patients with skin diseases, clinical strains of the fungi of the genus Malassezia. The aim: improving the efficiency of laboratory diagnostics malaseziosis on the basis of in-depth study of the biological properties (microscopic, culture, biochemical, genetic, antigenic) clinically relevant strains of the fungi of the genus Malassezia, circulating in Ukraine. Methods of research: microbiological, molecular-genetic, immunology, bioinformation analysis, statistics. Equipment: microscope, centrifuge, immunosorbent analyzer, thermostat, autoclave, PCR thermal cyclers, spectrophotometer, automatic dispenser, laboratory scales. For the first time in Ukraine, the species spectrum and prevalence of fungi of the genus Malassezia are determined in patients with dermatoses. The primary structure of the ITS of the rRNA gene of six strains of the fungi of the genus Malassezia was determined, which allowed the bioinformation analysis to perform their exact phylogenetic identification. For the first time in Ukraine, a diagnostic antigen was obtained from the M. sympodialis 97 strain, the primary structure of the ITS rRNA gene differs by 1% from the typical M. sympodialis strain ATCC 42132. On the clinical strains Malassezia spp. in experiments in vitro, a MIC liposomal-fat form of terbinafine was determined, which is 10 times more effective than the MIC of the terbinafine substance. Methods of isolation, cultivation, typing of the fungi of the genus Malassezia and determining their sensitivity to antimycotics have been developed and introduction into practice. A method for diagnosing mycosis of feet, brushes and onychomycoses based on PCR diagnostics using the panfungals primers ITS 4 and ITS 5 was developed (patent of Ukraine № 68645). A strain of M. sympodialis 97 is isolated and characterized, which is the producer of a specific antigen for the manufacture of test systems for the diagnosis of malasseziosis skin diseases, including atopic dermatitis (patent of Ukraine № 93898). The isolated strains of Malassezia spp.for the first time in Ukraine have been deposited in the museum of microorganisms pathogenic for humans. The degree of implementation: patents and acts of introduction of teaching programmers of the medical universities of Ukraine and of the research establishment. Scope (area) of application: medicine (microbiology, dermatology).