Study On A Dot Immuno-gold Filtration Assay For The Detection Of Porcine Parvovirus Antigen

Porcine parvovirus (PPV) was proliferated on PK-15 cell by the way of synchronous inoclation, then cultured PPV were purified and concentrated with contrifuation and PEG-6000, the volume of which was decreased by fifty-two times and the content of virus protein was 1.5mg/ml. After the oil emulsion antigen prepared with the purified and concentrated PPV inoculated into the rabbits for several times, the high immunse sera were obtained, IgG from rabbit anti-PPV serum was extracted by salting out, gel filtration, dialysis and so on. The purified rabbit anti-PPV IgG was labelled with the prepared colloidal gold. Dot-immunogold filtration assay (DIGFA) with the Nitrocellulose Membranes as carrier and the red colloidal gold as label was elementarily developed for detection of PPV and the optimum working conditions were determined. The minimum amount of PPV antigen detected by the DIGFA was 5.86ng per dot, which was 100-fold as sensitive as the haemagglutination(HA) test.Specific assay showed that DIGFA didn't happened cross-reaction with PRRSV, PRV, PK-15 cell culture media, HCV tissue, naturel porcine liver and kidey.