Establishment And Preliminary Application Of Indirect ELISA Method For CSFV Antibody

This study was conducted by taking E2 protein as an Antigen to be coated, groping the required conditions for establishing an ELISA. The results show:optimal coating concentration of 40ng/hole; the detected serum was diluted by 1/100 and incubated for 30 min; 1/600 diluted HRP secondary antibody was chosen to react for 30min; substrate effected for 15min before a reading of OD450. The sensitivity of the indirect ELISA for CSFV antibody established in these conditions is 80.5%.the specificity is 82.5%. and the coincidence rate was 80.6% with IDEXX Kit.The test results of the classical swine fever virus antibodies of immune-tracking serum in 8 pigs show that:being vaccinated in 1 copy/head, the whole CSFV antibody levels of the piglets reached the peak after a vaccine immunization for 60 days, and the antibody levels can be maintained at a high level of 105 days or more.The test results of the classical swine fever virus antibodies of immune-tracking serum in I and II farms show that:this indirect ELISA for CSFV antibody established in these conditions has a better consistency with IDEXX Kit.80 sows in I farm wholly have a high antibody level and a good uniformity. The whole antibody level of pigs has improved until 50 days after immunization. The established indirect ELISA for CSFV antibody is better than IDEXX Kit at the distinction for high antibody levels in serum samples. On the Ⅱ pig herd immunity day.14 pigs’antibodies were negative level, which indicates that the first immunization time is unreasonable.33 days after the first immunization, antibody levels had not all turn positive, which would lead to a longer window period of whole herd. Comparing the established indirect ELISA for CSFV antibody with IDEXX Kit, the former can detect antibody earlier, being conducive to detect antibody levels after an immunization of hog cholera vaccine for pigs.