To Built The Diagnosis System For The Porcine Circovirus By The Immuohistochemistry Assay

To build the diagnosis system for the porcine circovirus type 2 by the two steps of immnuohistochemistry assay(Envision assay),the plasmid of PGEX-ORF2 was transformed to E.coli BL21,and PCV-ORF2 fusion protein was expressed after induction by IPTG.Prepared anti-PCV2-ORF2 antibody and choosen the best working concentration,then the rabbits were immunized by the renaturated of PCV-ORF2 fusion protein.Used the two steps of immunohistochemistry assay to detect the lung of Porcine circovirus that doubtful illness tiers in the clinical,considered the anti-PCV2-ORF2 antibody was the first IgG,and then used the diagnosis assay of RT-PCR and an indirect enzyme-linked immunosorbent(iELISA)assay to confirm the consequence.The results of the two steps of immnuohistochemistry assay(Envision assay)directed exactly from localization,qualitation and quantitation,so that completed the standard of the diagnosis system.The experiment consequence certificated that the purity and the specificity of PCV-ORF2 fusion protein were very hight,the titer of the antiserum obtained in this experiment was up to 1:10240(2¹⁰).The masculine cell of lung tissue where existed in the intracytoplasm from the porcine alveolar macrophage,alveolar epithilal cell and bronch epithelium mucosae.The diagnosis system of immunohisochemistry assay did not only use in the laboratory differential diagnosis but also used in the basement layer veterinary surgeon generally, it identified the diagnostic antigen in time and improved the rate of clinical definite,it prevented the disease and reduced the encomical loss effectually.The research offered the new ideas and new methods for the other animals viral disease.