Analysis And Application Of African Swine Fever Virus P30 And P54 Protein Antigenic Epitope

African swine fever(ASF)is an acute,vigorous,highly dead infectious disease of pigs caused by African swine fever virus(ASFV).ASF first broke out in China in August 2018,dealing a heavy blow to China's pig industry.The traditional antigenic epitope screen is high,and the cost is high.With the development of bioinformatics,the computer can be screened by the sequence of gene sequences in accordance with different algorithms,which is widely used in its convenience and accuracy.This study uses biological information to obtain 10 advantageous antigenic epitopes based on the main structural proteins p30 and p54 of ASFV.It is predicted to obtain 10 advantageous antigenic epitopes in the flexible Linker to the p ET-28 a expression vector,expressed by the prokaryotic expression system,purification Recombinant multi-epitope antigen.The electrophoretic gel results show that the recombinant protein is about 24 KD,which shows that the protein can be reacted with the labeled antibody and the ASFV positive serum,and the recombinant protein is immunized to mice,and the flow cytometry is used in phenotype CD3+,CD4+,CD8+ fluorescent staining,and CCK8 proliferation were significantly higher than that of the control group.How to enhance cellular immune response and produce high levels of neutralizing antibodies are key issues of ASFV vaccine research.Therefore,this study was prepared based on dendritic cells(DC)-based bionic nanoparticles.After separating the bone marrow source from the mouse,the fluorescent labeling CD11 C antibody After the purity was detected,the induced DC was stimulated using a multi-epitope antigen,and the DC cell membrane containing antigenic information was separated on nanoparticles,nanoparticles.It is obtained by dispersing and will induce the IL-2 package that can induce CTL responses by PLGA nanomaterials.Thereby obtaining bionic nanoparticles having an antigen with a double function of the cytokine.The particle size analyzer showed that the nanoparticle PLGA-NP size was about 170 nm,and the bionic nanoparticle diameter was about 190 nm,and the promiscuous nanoparticles were observed to have a significant nuclear membrane structure and electrophoretic gel demonstrates the complete DC membrane protein on bionic nanoparticles.Analysis of experimental identification of bionic nanoparticles Western Blot using CD11C?CD86 antibody,the results prove that key membrane proteins have reactive principles.The author uses the purified multi-surface recombinant protein as a coated antigen,establishes an antibody detection method of ASFV,and uses the checkerboard titration to determine the optimal reaction conditions and the negative serum sample determines the critical value of the Cut-OFF,and determine a large number of known backgrounds.Serum samples were established ROC curve analysis,which proves that 98.72%(95% CI,93.06 to 99.97)were obtained at a critical value of 0.338,and the diagnostic specificity of 98.14%(95% CI,95.31 to 99.49)was proved.The positive serum of other pigs,such as foot-and-mouth disease,swine fever,and pigs,etc.,there is no cross-reactions.In order to explore the selection reactivity in the multi-epitope recombinant protein,the epitope synthesis peptide segment was detected by an ELISA method,which demonstrated that two epitopes and ASFV positive serum reactivity in part p54 were significantly higher than other groups.In summary,this study obtains the ASFV protein p30,p54 antigen epitope by predicting the screen,which is intended as a multi-epitope antigen,and the prokaryotic expression system purifies recombinant protein,and immunological experiments are carried out,and the recombinant protein is based.The multiepisode bionic nanoparticles were preliminarily studied,and the detection method of antibody was established.It is of great significance for the research of the ASFV vaccine and diagnosis.