Development and testing of a test system for polymerase chain reaction in order to detect alpha-toxin <i>Clostridium Perfringens</i>

Abstract

With the spread of clostridiosis, epidemiological and epizootic safety is violated, the degree of which depends on the immunological status of the animal, therapeutic and preventive measures in the livestock complex, the toxigenicity of the bacterium and the severity of the disease. The Clostridium perfringens bacterium is ubiquitous, produces 20 different toxins and pathogenicity factors, one of which is alpha-toxin, which is found in meat and other biological samples, which threatens the biosafety of livestock enterprises, markets, shops, etc. The article is devoted to the development and testing of test- systems for real-time polymerase chain reaction (PCR) to detect the DNA of the gene encoding the alpha-toxin of the Clostridium perfringens CP322 strain, which is widespread in the Russian Federation. The synthesis of oligonucleotides proposed by a team of scientists from Korea Chon J.W., Park J.S., Hyeon J.Y., Park C., Song K.Y., Hong K.W., Hwang I.G., Kwak H.S., Seo K.H. in 2012 served as the basis for the creation and testing of the test system [1].Optimization of the diagnostic technique, selection of the correct concentrations of reagents and DNA in samples from cattle, synthesis of highly specific primers and modernization of the probe molecule formed the basis for the development of a test system, which was tested on 319 samples - biomaterial obtained from cattle. The proposed method of molecular genetic diagnostics will allow veterinarians to timely identify the causative agent of enterotoxemia in the test samples, make a comprehensive diagnosis based on epizootic, clinical, pathoanatomical and laboratory data, prescribe effective treatment and develop an action plan to prevent the development and spread of infection.