Establishment And The Preliminary Application Of The Indirect ELISA Of Examining The Antibody Of GPV

Gosling Plague is a violent disease which is caused by Goose Parvovirus as a member ofthe Parvovirus genus of the Parvoviridae. It mainly threaten one-month goose and muscovyduck and induce an acute, highly contagious and highly mortal disease. The threat to animalscorrelate close to age. The morbidity and mortality in 10 day age reach to 100% in infectiousherds. Infectious rate decrease with the age increase. But some reports said there is aincreasing tendency of infectious age in recent years. Infectious rate went up to 14.3% over 30day. This may be caused by enhanced virulence and genovariation. GPV was first found inchina by fang dingyi in 1956 and then was reported all over world and brought a great threatto geese.Our experiment aim to establish an indirect ELISA to detect the antibody of Gooseparvovirus and applies initially in the field trial. The goose parvovirus generally does notcause the morbidity to big goose, but may cause the horizontal transmission in big goose withthe silent infection, continuous holding and expelling virus. This method not only detectantibody in serum of healthy goose but also detect the immunized especially to the silentinfectious goose. There is an important significance in decreasing horizontal and verticaltransmission. So far domestic and foreign has reported that detecting antibody of gooseparvovirus by agar diffusion reaction and the neutralization experiment, but some scholarsreported the sensitivity of the AGID was low, and needed high-titer antiserum and theconcentrated allantoic fluid, the neutralization experiment was hard to adopt in practicalapplication because of operation-tedious and time-consuming. In order to make up thedeficiency for the two methods above we establish the ELISA to detect the antibody of GPVand make a substructure for detecting the level of antibody and epidemiologic survey.Antigen was proliferated with Duck embryo in the experiment, then purified bychloroform processing, differential centrifugation, PEG precipitates and Sephrose-4B columnchromatographic. And then prepared high—immunization serum as the reference ofmasculine serum of ELISA by our own and the purchased antigen. The OD value in detectionexceeded 1.0. The reference of negative serum of ELISA were collected 50 nonimmunizedgeese which were around 20 day from non-epidemic countryside and the OD value was below 0.3. Purified IgG from geese serum and immunized rabbits to prepare enzyme-antibody whichof the titer was 1: 6400. we Groped the condition of reaction,established ELISA and definedthe critical value of the experiment is 0.35.Preliminary application in Field trial we detected436 big geese and the masculine rate of GPV antibody was 87.49% and the negative rate of239 gosling was 94.08%.Through the experimental, we has established indirect ELISA and replenished thedomestic blank. The method is a sensitive, operation conveniently and fiting for application inpractice also in laboratory diagnosis.