The Theoretical And Practical Aspects Of The Recombinant Antigens For The Diagnosis Of Bovine Leukemia Production

O.V. Shapovalova
2016 Zenodo  
Introduction. Nowadays the problem of bovine leukemia (EBL) effective diagnosis in countries where EBL is registered and the disease-free areas remains actual. The main diagnostic tests are immunodiffusion reaction (AGID) and enzyme-linked immunosorbent assay (ELISA), which allow the identification of infected animals by the presence of antibodies to bovine leukemia virus (BLV) both in serum and in milk samples. The effectiveness of these methods depends on the quality of diagnostic test
more » ... used and determines by the cultural and recombinant virus antigens specificity. EBL recombinant antigens have certain advantages as they are more active and cheap. Purpose of the work. The analysis of theoretical and practical approaches in the bovine leukemia virus recombinant antigens development and its diagnostic potential evaluation. The article contains data from the literature on the recombinant antigens of bovine leukemia virus construction and use. Analysis of the literature showed that the recombinant proteins are widely used in the serological diagnosis of bovine leukemia. Numerous protocols of BLV gp51 and p24 immunodominant antigens preparation has been developed in heterologous systems (Saccharomyces cerevisiae, E. coli, vaccinia virus, baculovirus). In order to obtain recombinant antigens, the BLV provirus genome regions isolated from FLK-BLV cell culture, lymphocytes or tumor cells from naturally infected cattle are typically used. For the recombinant antigens labeled by hexahistidine or Srept II purification one-step immobilized-metal affinity chromatography IMAC and highly selective Strep-Tactin affinity chromatography methods are carried out. The end products activity and specificity are studied in the immunoblotting, ELISA and AGID diagnostic reactions. The ukrainian scientists' publications are devoted to the clone E. coli HB101-2 transformed by the recombinant plasmid containing fully functional BLV env and gag genes nucleotide sequence construction. The efficiency of BLV gp51 and p24 encoding re [...]
doi:10.5281/zenodo.192322 fatcat:qevi5mnh6vc7xfjxg5ivvmamyu