The Application Of Neospora Caninum Nc SRS2 Recombinant Antigen In Diagnosis And Immunoprophylaxis

Neosporosis and Toxoplasmosis are reproductive barrier parasites diseases,respectively caused by Neospora caninum and Toxoplasma gondii parasitize a nuclear intracellular of mammals,they will lead to non-pregnancy,miscarriage,stillbirth,malformation of pregnancy females and newborn developmental delays.There is no effective drugs and vaccines to preventing this disease yet,control is the key to early detection,identify diagnosis,adopt comprehensive prevente and control.Therefore,this paper aimed to Neospora caninum NcSRS2 study of the application of recombinant antigen protein,on one hand,by the "female abortion syndrome" to make rapid detection and diagnosis of the difficult disease,on the other hand for Neospora caninum vaccine research and development to lay the foundation.(1)According to Neospora caninum NcSRS2 recombinant plasmid,recombinant antigen and positive serum as the research material,optimizing expression,purification methods,the laboratory has saved plasmids,strains and recombinant protein is activated,optimization and standardization,to explore Neospora caninum NcSRS2 surface antigen detection characteristics of different stages,strain energy and storage life.Results show that save the species has the deactivate from 2009 to 2012;Saving the recombinant protein has been gradually degraded,and is preserving strains has good vitality from 2013 to 2015,after recovery,activation can continue to protect bacteria and the subsequent test.Strains and 30%glycerin preserved in a certain proportion of-20? for 2 years.(2)Applying the team to establish rELISA antibody detect method,the collected from3592 samples of 13 counties and cities of horses,cattle,sheep serological detection.According to the results of the Neospora caninum average positive rate of 14.1%(95%CI:13.0~15.3),Toxoplasma gondii the average positive rate was 13.5%(95%CI: 12.4~14.6),and the mixed infection rate was 0.6%(95%CI: 0.3~0.9).Results of different ages 1 to 3 years old more susceptible.It is common in the plant-eating livestock,is the main source of female abortion.The region's horse infected Neospora caninum and Toxoplasma gondii is reported for the first time.(3)Using Neospora caninum NcSRS2,NcSAG1 recombinant protein after mixed with Freund's adjuvant to preparate recombinant protein to immune BALB/c mice,using indirect ELISA method were determined IgG,IgG,IgG2 a,IL-4 and IFN-? expression level.The results indicated these could elicit both Th1 immune response with elevated levels of IgG,IgG2 a and IFN-?,and Th2 immune response with elevated levels of IgG1 and IL-4.Moreover,the levels of IgG were also elevated.Antibody levels be found that twice immune very significant in adjuvant group and PBS control group(P<0.01),and the effect of recombinant protein NcSRS2 were significant in NcSAG1 recombinant protein and comprehensive immune group(P<0.05).