The Development Of Colloidal Gold Immunochromatographic Assay For Detecting Antibodies To Canine Parvovirus-2

In this study, a fast, sensitive and specific method was developed for detectingantibody to canine parvovirus-2 with the colloidal gold immunochromatographic assay.In this experiment, we used F81 cell line to canine parvovirus-2(CPV-2) wasisolated from faeces specimens of dogs which had seemingly been infected by CPV-2,F81 cell line cultured which susceptible to CPV-2 and then, immunized dogs to procuringthe positive serum anti-CPV-2; Collected serum of healthy dogs, purification of dog IgGwas purified by the methods of (NH₄)₂SO₄ salting-out and Sephadex G-200chromatography method, and a high purity coefficient of IgG was proved by SDS-PAGEelectrophoresis method; Using canine IgG to immunize rabbits on common practicemothed, the antiserum was detected by double agar diffusion method, collecting rabbitsserum when the titer was above 1:64, the rabbit IgG was purified by caprylicacid/ammonium sulfamate salting-out method. a high purity coefficient of rabbit IgG wasproved by SDS-PAGE electrophoresis method. Synthesize 20nm colloidal gold particlesby trisodium citrate disoxidation method, rabbit anti-dog IgG was labeled with the goldsol, the optimized amount and pH value of labelling were 10μg/ml and 9.0, respectively.The labelled rabbit anti-dog IgG was purified on low temperature ultra centrifuging. Thepurified labelled antibody was diluted at ratio of 1:2 and adsorption on fiberglass. The testlines was CPV-2 striked on the nitrocellulose filter, concentration of CPV-2 was 0.8mg/ml.The control line was goat anti-rabbit IgG which concentration was 0.75mg/ml strked onthe nitrocellulose filter.Used the dipstick to detect the serum anti-CPV-2, the result was gained in tenminutes, when the titer of hemagglutination inhibition test was higher than 1:80, thecoincidence of colloidal gold immunochromatographic assay and hemagglutinationinhibition test was 100ï¼…, the result proved colloidal gold immunochromatographic assaycould substitute hemagglutination inhibition test to detect the effect of immunity toCPV-2. Colloidal gold immunochromatographic assay detected the serum anti-CPV-2, theoperation was simple and the short time was needed, provided a useful tool for thedetection of serum anti-CPV-2 titer.