Development of a method for obtaining bovine adenovirus subgroup 1 antigen

Relevance. Adenoviral infection of cattle remains one of the main problems of modern dairy and beef cattle breeding. The effectiveness of health measures in intensive livestock farming depends on the timely detection of infected livestock, which in turn is determined by the serological activity and specificity of the antigens used in diagnostic kits.

The aim of the study was to develop a method for purification of adenovirus antigen of the first subgroup of adenovirus infection in cattle (cattle).

Methods. The serological activity of the components of the causative agent of adenovirus infection in cattle was studied using the immunoblotting method and enzyme immunoassay. Obtaining the bovine adenovirus antigen included the preparation of a bovine adenovirus mat brood on ВНK-21/13, the production of viral biomass by roller cultivation, the destruction of infected cells by ultrasound and the release of cellular fragments by centrifugation, precipitation of the antigen using PEG-6000, and concentration of the virus through a step gradient of cesium chloride by ultracentrifugation.

Results. As a result of separation of the viral material in a stepwise density gradient of cesium chloride, three antigenic fractions were obtained. The most purified was fraction № 1, which contained a major region with a molecular weight of 50.0 kDa and showed maximum antigenic activity. The antibody titer detected in the blood serum of rabbits on the 45th day after their immunization with this fraction was 1:3200. The results obtained open up prospects for the use of the adenovirus antigen of the first subgroup purified by the authors in the design of a test system for screening studies for adenovirus infection in cattle and monitoring the effectiveness of antiepizootic measures.

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