Establishment of ELISA method to detect porcine circovirus 2 antibody.

The research established a fast and sensitive enzyme-linked immunosorbent assay(ELISA) method to detect porcine circovirus Ⅱ(PCV2) antibody.The experiment sieved four PCV2 antigens from differential centrifugation,gluco-discontinuous gradient centrifugation, cesium chloride isopycnic gradient centrifugation and CHCl3 extract bonding ultracentrifugation.We determined PCV2 antigen from CHCl3 extract bonding ultracentrifugation was the best antigen.In addition,we also confirmed the optimal coating-antigen concentration,HRP-SPA concentration,test serum dilution,blocking liquid concentration,blocking time,HRP-SPA reaction time,critical value,specificness,stability,accordant rate by ELISA method.The results revealed that for the positive serums of pig′s several viruses detected by this ELISA method,only the positive serum of PCV2 was positive;The six serums were detected four duplicates by this ELISA method,F test presented P0.05,the variation was quiet;The coincidence was 88.89% with "INGEZIM" PCV2-ELISA Kit.The results indicated the optimized ELISA was a specific,sensitive and stabile method,may detect PCV2 antibody.