The use of a synthetic peptide based on the immunogenic epitope GP51 for the serological diagnosis of bovine leukemia virus

Diagnosis of leukemia remains one of the key problems in carrying out recreational activities in farms that are disadvantaged by this disease. In the Russian Federation, various methods are used to identify the causative agent of leukemia, including serological methods, one of which is enzyme immunoassay (ELISA). The principle of the indirect variant of ELISA is to identify a complex formed by an antigen immobilized on the surface of the wells of a polystyrene tablet and specific antibodies contained in blood serum samples or cattle milk. The present work is devoted to the study of the possibility of using a synthetic peptide, which is an amino acid sequence of epitopes of the gp51 protein of the leukemia virus, as an antigen for ELISA in the indication of the bovine leukemia virus. To develop the ELISA method, we designed and synthesized the antigen – synthetic peptide BLVPEP (NCKYSNQCGDQGSFYVNHQILFLHLK QCHGIFTLTWEIWNC). In order to best sensitize this peptide, the main parameters of the ELISA formulation were worked out. In particular, the optimal concentration of the peptide for sensitization of the tablet was determined - 1.0 micrograms/ml, blocking solution – 3% solution of skimmed milk powder, anti–species peroxidase conjugate - 1:40000, the reaction buffer of the interaction "antigen-antibody" - FSB-T (pH 7.3) was selected, the minimum titer of antibodies to the pathogen, detectable in the blood sera of cows with leukemia – 1:1600. The ELISA method presented in this paper can be used as a basis for creating a test system for detecting the causative agent of bovine leukemia and find wide application in veterinary practice for serological diagnostics.

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