DEVELOPMENT OF PCR KITS FOR BACTERIA OF THE GENUS CLOSTRIDIUM AND TOXINOTYPES CL. PERFRINGENS IDENTIFICATION

Abstract

Bovine clostridiosis is one of the major problems in veterinary medicine, animal husbandry and food safety. Clostridia cause zoonotic infectious diseases. The causative agents of clostridiosis produce highly resistant spores that can persist in the environment for a long period of time. They are also present in the gastrointestinal tract and as spores in the tissues of healthy animals and can cause disease due to temperature or productive stress, mineral imbalances, injury and impaired immune response. The aim of our work was to develop new and optimize existing PCR protocols for the diagnosis of Clostridium bacteria and Cl. perfringens toxinotypes. The relevance of the development of PCR kits is associated with the lack of commercially available ones for the diagnosis of many types of clostridia. This makes it difficult to make the correct diagnosis and select rational therapy and vaccination. Materials and methods. Developed and optimized methods for the diagnosis of clostridiosis are based on real-time PCR and conventional PCR. This article presents the results of the development of PCR methods for rapid and inexpensive diagnosis of clostridial infections that are significant for livestock farming. For the first time, PCR kits were created for the detection of Cl. sordellii, Cl. novyi (types A, B) using conventional PCR and Cl. histolyticum in real time. Moreover, we have optimized PCR conditions and reagent mix for the detection of Cl. chauvoei, Cl. septicum in real time, Cl. tetani, Cl. botulinum using conventional PCR as well as a multiplex PCR for Cl. perfringens (A-E) typing.

Keywords

• pcr
• clostridium
• clostridial infection
• anaerobes
• real-time pcr
• electrophoresis
• cattle