Real-time PCR for indication of representatives of <i>Chlamydiales</i> order: needs for development and main analytical characteristics of the method

Abstract

   Respiratory diseases occupy a dominant position in the structure of morbidity, often the etiology of which remains unclear. This situation is largely due to the lack of knowledge of the entire spectrum of pathogens that cause respiratory diseases, as well as the lack of approaches to their indication and identification. Thanks to constant progress of molecular biology, a number of bacteria related to chlamydia have been identified, some of which are associated with human respiratory diseases, including chlamydia-like microorganisms. The role of atypical pathogens such as chlamydia-like microorganisms Parachlamydia acanthamoebae, and Simkania negevensis in the pathology of the human respiratory tract is still not fully understood. A real-time PCR-based method has been developed and optimized. It is capable of detecting the DNA of both chlamydia and chlamydia-like microorganisms (Parachlamydia acanthamoebae and Simkania negevensis). The method is characterized by high specificity and reproducibility, as well as by analytical sensitivity at a level of 8.7∙103 GE/ml. Its use will make it possible to assess the contribution of these atypical pathogens to the etiological structure of infectious diseases of the respiratory tract.