The Preparation Of Monoclonal Antibody To P1 Protein Of Canine Distemper Virus And Its Preliminary Application

Canine distemper (CD) is a highly contagious and fatal disease of dogs caused by the canine distemper virus(CDV).CDV vaccines have been widely adopted,but the hosts is becoming wider and the stain is becoming stronger.so get a right diagnosis is an important task.P genes are 1655 ribose nucleotides in the genome of the 1742 ~ 3396 nucleotides,which consists of two partially overlapping P gene reading frame encoding three proteins, structural proteins P, non-structural protein V and C. CDV P protein is a structural protein in the second protein with polymerase function. The virus is detected as phosphorylation of P protein 12h after inoculation, which emerge early and last during infection. P genes in the virus's lineage are extremely conservative.So choose it as a diagnostic antigen or antibody preparation is feasible. We first expressed and purification P1 proein in china,by IFA we found P1 exist in nucleus of infected cells,that this protein has connection with recomplication.Then we immuned SPF BABL/c with P1 protein and prepared monoclonal anitibodies ,we ge two strains, IgG2b substye.Make rabbit mutianibodies using purified CDV .Purified antibodies by ProteinG pillar .On one hand ,we made rabbit serum colloidal gold probe and made two stain monoclonal antibodies as detect line,made goat anti-rabbit IgG as control line ,then we got Gold Immunochromatographic assay(GICA),which could detect CDV 1X105.4 TCTD50/ml cell cdv ;on the other hand,we got two monoclonal anitibodies coating ELISA board ,bio-bind mutiply antibodies which can detect virus 5X103.4 TCTD50/ml.