Creation of a hybridization probe for the diagnosis of classical swine fever by the FISH method

Abstract

The article presents data creation of the DNA probe labeled with fluorescein-12-dUTP for its subsequent use in in-situ hybridization in cryotomic histological sections from samples of internal organs of pigs. This diagnostic method is especially relevant for the inapparate course of infection, which contributes to maintaining the viral load inside the herd without showing pronounced clinical signs. In addition, the method helps to identify the viral genetic material in the pathological samples during the post-viremia period. In this article the stages of creating a probe consisting of 173 pairs of nucleotide bases corresponding to the site of the glycoprotein E2 gene of the classical swine fever virus are shown. The final DNA probe was tested on tonsils from pigs infected with the CSF virus as a positive control for the DNA probe and as a negative control for the DNA probe is tonsils from healthy pigs not vaccinated against CSF were used. Finally, we obtained positive results of using the DNA probe, reflecting its high specificity and selectivity in the studied samples of positive control. The data obtained by us formed the basis for our further research in the field of infectious pathology of farm animals.