Immunizing Mice using Recombinant Truncated p72 Protein of African Swine Fever Virus and Establishment of an Indirect ELISA

摘要

African swine fever (ASF) is a serious infectious pestilence characterized by bleeding in domestic pigs. Therefore, it is necessary to develop effective methods to diagnose this virus, serological detection of specific antibodies against ASFV infection is important for successful clinical diagnosis. In this study, E. coli was used to express the truncated P72 (tP72) gene cloned into the prokaryotic expression vector pET28a (+). Rosetta (DE3). An indirect ELISA assay which against African swine fever virus (ASFV) was established by using purification of recombinant tP72 protein as coated material for detection antibodies. Most effective in exhibiting positive result was observed when the coated material at a concentration of 3.625 μg/mL, serum was diluted to 1:160 and the concentration of HRP-conjugated secondary antibody was 1:2000. Our results showed that the method displayed an excellent specificity (100%) and better sensitivity (1:1600) during serological test based on the criterion of an average value plus three standard deviations.