NANOTECHNOLOGICAL APPROACHES ТО STUDYING CHLAMYDIA TRACHOMATIS STRUCTURAL ORGANIZATION AND PATHOGEN DETECTION
Objective. Development of nanotechnological approaches for studying the structural organization of Chlamydia trachomatis and detection the pathogen using functionalized nanoparticles were the purpose of the study. Materials and methods. We used the C. trachomatis strain (serovar D). Cultivation was carried out on the McCoy cell line (infection titer was 5.5 gCPDJml). Identification of the pathogen was performed using PCR, fluorescent antibodies assay and cytological method (staining by Romanowsky—Giemsa). For C. trachomatis local immobilization microstructured sensor coatings of heparin and polyclonal antichlamydial IgG were created using microcontact printing. The pathogen was visualized in the atomic force microscopy contact and intermittent-contact modes. In addition the possibility to use fluorescent nanoparticles conjugated with polyclonal antichlamydial IgG for C. trachomatis detection in samples of biological material was studied. Results. Micro sensor coatings were made of polyclonal antichlamydial IgG and heparin allowing immobilization of various extracellular morphotypes of C. trachomatis. AFM analysis showed heterogeneity not only in the particle morphology and size but also in the surface antigens of C. trachomatis EB structural organization. Chlamydia mature and ultrasmall forms were detected and described. A specific pathogen morphotypes glow signal was demonstrated when processed by functionally-active nanoparticles allowing elaboration of a new generation of test systems based on nanotechnology approaches. Conclusion. The developed nanotechnology-based approaches can be used not only for C. trachomatis detecting in biological samples but for visualizing and studying the pathogen morphology, too. It has been shown that the C. trachomatis extracellular forms various morphotypes structural organization surfaces are heterogenic. All of the particles morphotypes contain antigenic determinants specifically interacting with polyclonal antichlamydial IgG, heparin and giving an intense fluorescence stable signal when processed by functionally active nanoparticles.