A Preliminary Study On Detection Method Of Dengue Virus Nonstructural 1 Antigen

OBJECTIVEPreparation of polyclonal antibody and Monoclonal antibody against DV(Dengue virus)nonstructural 1 protein and study on its specificity, establishing a primary method of DV nonstructural 1 antigen detection.METHODS1. The full-length DENV-NS1 gene was expressed with Pichia Pastoris expression system. Anti-NS1 polyclonal antibody was obtained after inoculating the BALB/C mice with the recombinant DENV-NS1 protein.2. Make research on the specificity of polyclonal antibody, using the methods of ELISA and indirect immunofluorescence(IFA).3. Recovery the Hybridoma Cell Line, Which is cultured and amplified to preparation mouse ascites antibody, using the methods of ELISA and indirect immuno fluorescence to study its specificity.4. Established a primary method of DV nonstructural 1 antigen detection, using indirect competitive ELISARESULTS1.After recovery, induced expression, identification, and low-temperature vacuum freeze-drying, we successfully got the purified DENV2-NS1 expressed protein powder.2. After the immunization of BALB/C mice with the expressed DENV2-NS1 protein, the high titer anti-DENV2-NS1 antiserum was obtained and the titer of the polyclonal antibody obtained in this experiment was up to 1:13500. we used ELISA and indirect immunofluorescence to confirm that the polyclonal antibody has cross reaction with the DV1,DV2,DV3,DV4.3.Successfull recovery the Hybridoma Cell Line, Which is cultured and amplified to preparation mouse ascites antibody, using the methods of ELISA and IFA to study its specificity, we can draw a conclusion.the NS1 monoclonal antibody has cross reaction with the DV1,DV2,DV3,DV4.4. The method of DV nonstructural 1 antigen detection was initially established.CONCLUSIONThe recombinant Dengue virus type 2 NS1 protein showed a strong immunogenicity. We successfully obtained the polyclonal antibody which has cross reaction with DV1, DV2, DV3, DV4. The Monoclonal antibody can bind NS1 have immune cross-reaction with DV1,DV2,DV3,DV4. We established a primary method of DV nonstructural 1 antigen detection.