By Dot-ppa-elisa For Rapid Detection Of Classical Swine Fever Antibody Methods To Establish And Expand The Application Of Research

A Dot-PPA-ELISA has been developed for the detection of antibody against Hog Cholera Virus (HCV) using HCV of itself as the antigen for the first time and the capability of it抯 relatively large scale application was determined. The technique was constructed mainly through following research steps-Comparison of different host system which is the best for the cell culture of virulent HCV and rabbit-adapted strain(C strain) of HCV, How to raise the titre of HCV in primary new-born piglet kidney cell and some factors which effect the multiplication of HCV C strain in the same cell line were studied, Comparison of some method for the detection of HCV C strain in cell culture liquid, The optimum conditions of Dot-PPA-ELISA was determined and the technique抯 application in an intensive pig farm was also carried out. The HCV C strain cultured in primary newborn big kidney cell was detected by the test of exaltation of Newcastle disease virus (END). Its purification degree was evaluated by SDS-PAGE, partly purified by centrifugation-polyethylene glycol precipitation- dialysis and the antigen had the characters of relatively high h purification degree, easy preparation, low cost. The method抯 positive standard as 1:60 was established by using the neutralization test in rabbits as control group. The 4 high specificity of the diaphragm which was made by dropping the antigen of HCV to the nitrocellulose membrane(as the solid phase carrier)was shown by the specific blocking test and the cross-reaction test ---it didn抰 react with the antibodies against 36 swine foot-mouth disease, swine bow-shaped body disease, porcine parvovirus disease, porcine pseudo-rabies disease (Aujeszky抯 disease), swine Brucella disease; the diaphragm had the ability of detection the positive antibody when it was diluted to 2.10 and so it has good sensitivity; stored at 4t at least for 9 months, at room tempeaure(10扖-.-25C) for 4 months, the diaphragm抯 sensitivity and specificity did not change and so it had good stability. The serum samples came from of growing pigs (160), sow (20), piglets that were newborn piglet immunized (160) were tested for antibody against HCV. The result was also evaluated and its feasibility in intensive pig farms was indicated. All the result have shown that Dot-PPA-ELISA was a convenient, rapid, sensitive specific useful method for the detection antibody and could be used for the surveillance of herd immunity and for the examination of immunization quality and so made provide for right proof to make rational immunity schedule.