Establishment Of An Indirect ELISA For The Detecting FPV Antibodies And Strains Screening Of Inactivated Vaccine

Feline parvovirus is called Feline panleukopenia virus again, the character of the infection disease chiefly originated from it are hyperpyrexia. and Vomitus. The grave decrease of white blood cell and inflammation. Under the nature condition, the FPV can infect many kinds of animal just like the felidae and the coon ursidae, such as tiger, panther, lion and raccoon,which has pose a serious threat to the economic animals and wildlife. But the minor type felid including the mink is the most easy one to be infected. The serosurvey showed that this disease can be found all over the world. And in our country the wild growed felid in different region all have the specific antibody against the FPV, which manifests that the FPV can exist and spread extensively among the wild growed felid under the nature circumstances. According to the dissection and the final diagnosis of the animal which was dead caused by disease, it can be seen that FPV is the main reason to lead to death and the tendency of the hazard to the felid and others is anabatic. However, there is not have any immune and therapeutic tool which is complete response to this disease in domestic at present, and there is not yet any vaccine which has the licence. It is evident that the current vaccines are not effective in protecting against infections with the genetically diverse field strains of FPV, and the attenuated vaccine viruses can revert genetically to cause clinical disease. So it has substantial significance to establish effective method to diagnose the FPV antibody and effective controlling measure.There are three expects to study in this research as follows: First, we'll prepare Rabbit anti-cat IgG /HRP and explore its preparative method. Now, Rabbit anti-cat IgG/HRP is rare in domestic market and they are very expensive. It is unfavorable for investigation and diagnostic treatment to spread and utilize. Therefore, investigate the preparative method of Rabbit anti-cat IgG/HRP in this experiment is necessary. First of all, it's should purify cat's IgG with octoic acid-ammonium sulphate and molecular sieve chromatography. Then it needs immunizated New Zealand rabbit adding Freund adjuvant. After that we have done Trialing bleeding with double agar diffusion and when potency reach up to 64 to begin to get blood and separation cony blood-serum. Then we have purified rabbit's IgG with octoic acid-ammonium sulphate and molecular sieve chromatography. Finally, the rabbit's IgG is linked with HRP using reforming odium iodate method. Besides we have shuded the approaching and purification method and got suitable technique line. Preliminary utilization also have proved the high performance of Rabbit anti-cat IgG/HRPSecondly, we want to establish an indirect ELISA method. Thus, in this experiment, indirect ELISA method of antibody test facility was established according to exploring and optimizing the condition such as level of virus antigen peridium, peridium condition, serum reaction time, level of Rabbit anti-cat IgG/HRP and reaction time, substrate action time, and so on. The method was demonstrated to be effective compared with the result of FPV hemagglutination inhibition experiment and established a new method for strains screening of inactivated vaccine. The data in this study demonstrated that the indirect ELISA was not only reliable with high sensitivity and specificity, but also was economical and convenient in terms of antigen production. Thus, it would be a very useful tool for routine diagnostics, epidemiological surveys, and outbreak investigations. Thirdly, we want to Screen strain of Inactivated Vaccine. Immunogenicity of vaccine is one of key factors to influence of immune effects, in addition, antibody regularity of growth and decline after animal immunization should be analysis. Therefore, strain of inactivated vaccine was screened by the indirect ELISA method and hemagglutination inhibition test to determine antibody level. Inactivators of superordinary are less effect immunogenicity of antigen. Adjuvant are able to strengthen immunogenicity of vaccine, this factors should also be studied. Lots of contents will be studied prospect, including animal origin source of antibody, regularity of antibody's growth and decline, protection test of vaccine, test of preservation of pure state time, field test and so on...