Application of IHC method for diagnosis of CSF

Abstract

The article presents experimental data on the reproduction of classical swine fever and the development of immunohistochemical method for the study of the obtained pathological material. 2 groups of animals were involved in the experiment (experimental infection group and non-control). During the experiment, blood was taken to obtain serum for further study, as well as organs for histological (paraffin embedded samples) examination and observation of organ pathology and native samples (frozen slices) for testing them in an immunohistochemical reaction. For indirect immunohistochemical reaction, primary mouse monoclonal antibodies specific to the recombinant protein E2 of the classical swine fever virus and secondary goat antibodies to mouse immunoglobulin labeled with horseradish peroxidase were used. Histological examination of the palatine tonsil, kidneys, spleen, liver, lung, lymph nodes and spinal cord brain revealed the development of pathological in all samples from the experimental infection group. The use of the ELISA method confirmed seroconversion of inoculated animals from the experimental infection group by the end of the study (DPI 14). Also, PCR studies have shown viremia in the same animals starting from the 2nd day of the experiment and up to the date of euthanasia of the animals at the end of the experiment, which corresponds to 14 days after infection. Despite the pathological changes in all the organs we studied, the indirect immunohistochemical method showed a positive result only in the spleen and in the palatine tonsil, while the number of positive sites was greater in the spleen. The nature of the chromogen distribution indicated the presence of the CSF virus antigen in the red pulp of the spleen, in leukocyte cells. The data obtained by us in the immunohistochemical reaction correlate with the data obtained in ELISA and PCR in the study of blood sera.