Zarichna O. Improvement of methods for detection of Mycobacterium tuberculosis and to study their sensitivity to new antibacterial compounds

Object of study: diagnosis of tuberculosis and antibacterial drug resistance. Objective: to trace the main morphological and cytochemical features of Mycobacterium cells, their sensitivity to new antibacterial compounds, improvement of methods for detection and identification of Mycobacterium tuberculosis. Methods: bacteriological, biochemical, electron microscopy, molecular genetic, epidemiological, statistical. A new semisynthetic nutrient media for culturing mycobacteria that are selective, differential-diagnostic, provide faster growth of mycobacteria, made of components available and cheap, easy to prepare were designed. Effectiveness of the method of PCR to detect the presence of Mycobacterium tuberculosis in clinical material of patients compared to traditional bacteriological methods was analyzed. The high antimikobakterial activity of new derivatives of fluorene on museum strains of Mycobacterium tuberculosis was established and usage of polyethylene glycol (PEG-400) to increase the solubility of compounds fluorenovoho number when performing bacteriological research was proposed. There was found that the studied compounds affect the reduction of enzymatic activity of Mycobacterium tuberculosis and atypical mycobacteria in vitro. There was found that in microorganism's cells under the influence of antibacterial compounds the ultrastructural changes were observed that indicate the violation of intracellular metabolism and inhibiting of the synthesis of cellular components. The results of microbiological studies of drug resistance of the strains of Mycobacterium tuberculosis to antimikobakterial drugs allow to predict future changes in these indicators and timely adjust the treatment regimen of patients and predict the effectiveness of therapy in inpatient phase of treatment. The environment proposed for further study in clinical laboratories for diagnosis of tuberculosis in research laboratories for culturing mycobacteria was developed. The combined usage of molecular genetic and culture methods, using our developed experimental semisynthetic nutrient media, greatly enhancing the efficiency of the detection and identification of Mycobacterium tuberculosis compared to using only one method. The compounds of domestic groups of fluoreniliden on mycobacteria there were the firstly synthesized; these substances were proposed for further pre-clinical studiing for determination of the possibility of their usage in practical public health and veterinary medicine. Scientific results obtained in the thesis on new methods of TB diagnosis and treatment of infections mikobakterial implemented in practice health health and used in the classroom.