The thesis is devoted to the development of diagnostics for identification and species differentiation of bacteria of the genus Chlamydia. For this we performed, bioinformational studies were carried out by aligning the primary nucleotide sequences of genes encoding 16S rRNA, RNase P RNA and MOMP of 10 chlamydia species. The level of homology and variability of the nucleotide sequences of the DNA of these three genes was studied, namely: each of 10 species of bacteria of the genus Chlamydia (species analysis); causative agents of chlamydia of farm animals, birds, carnivorous family of felines and rodents (grouped analysis) and all kinds of bacteria of the genus Chlamydia at the genetic level (general analysis). Designed for each of the 10 types of chlamydia, design of oligonucleotide primers for unique DNA regions of the gene, coding gene of the main membrane protein MOMP. Practically, the ratio of the reaction mixture and the amplification regime of the primer systems was worked out to obtaining clear bands of a given size on electrophoregrams of samples of control DNA of bacteria of the genus Chlamydia of all 10 species (C. abortus, C. avium, C. caviae, C. felis, C. gallinacea, C. muridarum, C. pecorum, C. pneumoniae, C. psittaci, C. suis). Optimal parameters of amplification at the annealing temperature of the primers are determined. Installed, that 10 pairs of oligonucleotide primers (direct and inverse) were constructed, are specific for the nucleotide sequences of the DNA fragments of each of the ten species of chlamydia and are not complementary to DNA sequences of pathogens, as other diseases of infectious etiology, and opportunistic microflora and prion proteins. The developed 10 PCR-methods for the indication and species differentiation of bacteria were tested for analytical specificity and sensitivity, as well as validation with the techniques underlying 7 other PCR-test-systems for the diagnosis of chlamydial infections. Determined analytical significance of developed diagnostics taking into account indicators specificity, sensitivity, accuracy and reproducibility. 35 samples of biological material were examined, namely: 11 samples of control DNA, 18 DNA samples of chlamydia from biological material from farm animals, 3 samples of DNA of leptospira, 3 samples of DNA of babesias. For holding the species differentiation, this or that chlamydial isolate, have been developed two algorithms for the application of these diagnostic PCR-methods, which make it possible to clearly identify the causative agents of chlamydia in animals. The aspects of the time difference are taken into account, labor-consuming PCR-methods of carrying out research on the identification of chlamydia and the costs of PCR reagents. Epizootic monitoring of chlamydial infection among cattle and pigs in farms of different regions of the Poltava region was carried out. Taking into account the results of screening studies, a specific circulation of bacteria of the genus Chlamydia was established. Diagnostic efficiency and significance of developed PCR-methods of diagnostics of polymerase chain reaction are established. The predicted value of positive and negative results of diagnostic significance is also determined. The scientific novelty of the constructed PCR-methods is confirmed by 10 patents of Ukraine on the utility model. The methodical recommendations for the use of the created system of testing the standard operating procedure have been developed. The results of the dissertation work were highlighted at scientific and practical conferences of different levels. Thus, the conducted studies allowed to design sufficiently sensitive, analytically specific methods for the indication and species differentiation of each of 10 species of bacteria of the genus Chlamydia by PCR.
