The Preparation Of Monoclonal Antibodies Against CSFV-C Strain And Development Of Rapid Test Strip For CSFV

Classical swine fever(CSF), a highly-infectious disease, brings severe economic losses to the pig farming worldwide.It has been the most important infectious disease in our county many years,and In order to control CSFV, many countries have paid a lot of manpower and money. In recent years, this disease showen many new characters.The incidence of this disease raised, the clinical representation became complex and the incidence of the immuned hogs raised. CSF is still the most important infection in our country.Monoclonal antibodies are produced by cell fusion technology. In contrast polyclonal antibodies. Monoclonal antibodies have following virtues: identical structure uniform component high specificity; animate biotic-activity and fewer crossing reactivity. Monoclonal antibodies of CSFV are prospective in this study, with the purpose of prevent and diagnose.To gain purified special mice anti-CSFV, six weeks old BALB/C mice were immunized. Antibody titers were detemined using indirect ELISA with CSFV in coating the micro-wells. The specific anti-CSFV sera were detected in all mice and were sufficient for subsequent experiment. Using monoclonal antibody technique with the immune mouse's spleen cell and cell's routine method of FO bone myeloma to merge with PEG1500, it is carried on for many times, the result is 328 in all merges wells.Screened by indirect ELISA, positive well that merged have 30 wells in all, selected 10 positive wells to carry on clone 3 times by diluted, and attaining one strains that stably excreted McAb hybridization tumour cell against CSFV-C strain. Using this tumour cell to produce ascitic fluid,the antibody of ascitic fluidv was detected by indirect ELISA.The titers was1:105and the titers was1:160 in the cell culture medium of hybridization tumour cell. Detecting the sample of ND,PPRS,TGEV and EDS-76 with indirect ELISA,the result showed no crossing reaction.A kind of method for determining swine fever virus (CSFV) by using colloidal gold immunochromatography assay (GICA) was developed. Using the reducing method of citric acid sodium prepare colloidal particles, marks and purify the colloidal gold particles with the CSF antibody. CSF antibody and goat-anti-rat antibody were fixed on nitrocellulose membrane and immunochromatography testing paper strip was manufactured for CSF diagnosis. Using the developed rapid diagnosis strip detect collected 70 treats examining sample, 26 positive cases were detected.While isolation and identification of virus have 28 cases of positive cases. The coincidence on positive rate of detection were 92.8%, diluted 5 positive CSFV sample, Perform the sensitivity experiments with testing paper stripe and ELISA method to 5 diluted positive CSFV sample.the results showed that the testing paper strip was higher sensitivity and had no cross reaction with ND, EDS-76,TGEV,PRRS. This diagnosis method was convenient, rapid and accurate.With high specificity, good repetition and without using any instrument. it was a great significance in the diagnosis and prevention of CSF.