Development Of McAb To GPV And The Study On ICS To Diagnose GPV

Gosling Plague was a kind of acute infectious disease that manly endangered young goose. It has a high mortality and severely threatens goose farm. Related study on the gosling plague was developed in the world and had got a certain achievement after the goose parvovirus (GPV) was isolated from the goose with goose embryo by Fang Ding-Yi in China in 1956. Because the diagnostic methods to the disease directly affect the development of goose farm, they were paid close attention to in the world..According to the method described in the paper of Li Mao-Xiang, 2.2% of NaCl and 6% of PEG were chosen as the conditions to sediment most of the virus, they were then purified through the Sepharose-4B chromatography. The purified antigen was obtained and was used to immunize 6 to 8 weeks old Balb/C mice. After four times immunization to the mouse, McAbs were prepared by fusing SP2/0 myeloma cells with spleen cells of Balb/C mice.78 hybridomas holes were observed, the fusing rate is 21%. 8 McAbs against to GPV were chosen by indirect ELISA. Positive hybridomas which could secreted McAb were cloned three times by technique of limiting dilutions, 3 kinds of McAbs were chosen, which were named 2B8,2B10 and 2H7. The golden immunochromatography reagent was developed by labeling the 2B8 McAb with colloidal gold and coating antibody horse anti-GPV on nitrocellulose membrane. A rapid immunochromatographic strip(ICS) for detecting GPV was established. The end (which was used to touch the sample) of the product was inserted sample liquid. The result can be observed in 5 minutes. When the red line only appeared in the control line, the result is negative. When the red line appeared in both control line and test line, the result is positive. Through groping the test conditions, the pH8.2 0.1% PEG-600-TBS was selected to dilute the McAb anti-GPV with colloidal gold (dilute it to l:4).The 0.01 M pH9.0 TBS was selected as the coating liquid(dilute the antibody horse anti-GPV to 1:4 and the antibody of sheep anti-mice to l:4).The 0.1%PEG-600-TBS was selected to the dilute sample.In the study, a rapid immunochromatographic strip(ICS) for detecting GPV were established. The diagnostic methods spare more time to diagnose the disease. It fits for the use of GPV diagnoses in the laboratory, as well as the use in the field.